Authorship：Last author   Language：Japanese   Publishing type：Research paper (scientific journal)  

COVID-19 became pandemic infection and medical schools in Japan were restricted medical education programs. In 2020, all lectures at Ehime University Graduate School of Medicine were canceled till April 22nd. Thereafter lectures were restarted using remote learning system. On June 11th, obligatory lectures and practices were begun as face-to-face classes keeping necessary social distancing. For example, large lecture rooms were used with reduced number of the students, and students were divided into two classes and teachers lectured the same contents two times. Bed side learning practice in the University Hospital restarted at the end of June, while practices in municipal hospitals were canceled. In the autumn semester 2020, face-to-face lecture and remote leaning were blended, and the faculty will care of infection prevention including students’ social distancing.

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Authorship：Last author   Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Ehime Medical Association  

(Objective) At 2017 and 2018, 20 medical schools involved Ehime University were evaluated by Japan Accreditation Council for Medical Education (JACME) using the World Federation for Medical Education (WFME) Global Standards for Quality Improvement of Basic Medical Education. We investigated differences of these medical schools.
(Method) Based on the evaluation reports written by JACME, we count partially fulfilled sub-areas, number of indications, high evaluations in these schools.
(Results)Ehime University has an average number of sub-areas partially fulfilled. The number of indications are higher especially basic standards. Among 20 medical schools, number of basic areas’ indications at area 1(Misson and outcomes) and other area are highly correlated.
(Conclusion)In WFME Basic standards, misson and outcomes of medical school is an important part.

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：愛媛大学教育・学生支援機構  

CiNii Books

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Other Link： http://id.ndl.go.jp/bib/030370522

Authorship：Last author   Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Ehime Medical Association  

（Objective）It is possible to hypothesize that if the graduation rate within 6 years of medical students is decreased, the pass rate of National examination is increased. But it is unknown that this hypothesis is true or not. <br />
(Method) Based on the data of National and public medical schools in 2016-2019, we investigated the correlation between a pass rate of National examination for medical practitioners and a graduation rate for six years. <br />
(Result) There is no or little relation between the factors used.<br />
(Conclusion) For increasing the pass rate, we have to search other methods to enhance students’ learning performance.

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To resolve the problem that evidence-based medicine (EBM) courses are not sufficiently taught in Japanese medical schools, we organized a year-round EBM-learning course. This study was an observational study and was designed to evaluate the participants' understanding of EBM using an original survey. The survey was given three times. In total, 18 students responded to our survey. Of those 18 students, six students answered both the first and the last surveys, and their mean score increased 1.17 of 4.00 (95% CI: 0.72-1.65). These results suggest our course improved students' ability to read clinical articles.

DOI： 10.1002/jgf2.38

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：BIOMED CENTRAL LTD  

Background: Physician-scientists play key roles in biomedical research across the globe, yet prior studies have found that it is increasingly difficult to recruit and retain physician-scientists in research careers. Access to quality research mentorship may help to ameliorate this problem in the U.S., but there is virtually no information on mentoring in academic medicine in Japan. We conducted a survey to determine the availability and quality of mentoring relationships for trainee physician-scientists in Japan.
Methods: We surveyed 1700 physician-scientists in post-graduate research training programs in 6 academic medical centers in Japan about mentorship characteristics, mentee perceptions of the mentoring relationship, and attitudes about career development.
Results: A total of 683 potential physician-scientist mentees completed the survey. Most reported that they had a departmental mentor (91%) with whom they met at least once a month; 48% reported that they were very satisfied with the mentoring available to them. Mentoring pairs were usually initiated by the mentor (85% of the time); respondents identified translational research skills (55%) and grant writing (50%) as unmet needs. Mentoring concerning long-term career planning was significantly associated with the intention to pursue research careers, however this was also identified by some mentees as an unmet need (35% desired assistance; 15% reported receiving it).
Conclusions: More emphasis and formal training in career mentorship may help to support Japanese physician-scientist mentees to develop a sense of self-efficacy to pursue and stay in research careers.

DOI： 10.1186/s12909-015-0333-2

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DOI： 10.11172/keitaikinou.11.17

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：VERSITA  

We investigated the effects of Rho-associated kinase (ROCK) on migration and cytoskeletal organization in primary human osteoblasts and Saos-2 human osteosarcoma cells. Both cell types were exposed to two different ROCK inhibitors, Y-27632 and HA-1077. In the improved motility assay used in the present study, Y-27632 and HA-1077 significantly increased the migration of both osteoblasts and osteosarcoma cells on plastic in a dose-dependent and reversible manner. Fluorescent images showed that cells of both types cultured with Y-27632 or HA-1077 exhibited a stellate appearance, with poor assembly of stress fibers and focal contacts. Western blotting showed that ROCK inhibitors reduced myosin light chain (MLC) phosphorylation within 5 min without affecting overall myosin light-chain protein levels. Inhibition of ROCK activity is thought to enhance the migration of human osteoblasts through reorganization of the actin cytoskeleton and regulation of myosin activity. ROCK inhibitors may be potentially useful as anabolic agents to enhance the biocompatibility of bone and joint prostheses.

DOI： 10.2478/s11658-011-0006-z

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Language：Japanese   Publishing type：Research paper (scientific journal)  

Formaldehyde or formalin is indispensable not only as a preservative but also as a disinfectant of cadavers for gross anatomy. It has recently attracted a great deal of attention as a health hazard for students and lecturers. To reduce the concentration of formaldehyde gas (FAG), we improved a novel local ventilation system of the push-pull type. This is the first report dealing with the effects of this ventilation system on the health of students before (over 1 ppm) and after (0.1 ppm) the installation. The percentages of students with lacrymal symptoms or airway irritation were reduced to a third of what they were before the installation. In particular, the number of those with continuously strong symptoms was reduced to a sixth of the pre-installation levels. This local ventilation system draws in fresh air from outside, and directs it to the breathing zone of the students, effectively reducing their symptoms.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

Intercellular adhesions between renal glomerular epithelial cells (also called podocytes) are necessary for the proper function of the glomerular filtration barrier. Although our knowledge of the molecular composition of podocyte cell-cell contact sites has greatly progressed, the underlying molecular mechanism regulating the formation of these cell-cell contacts remains largely unknown. We have used forskolin, an activator of adenylyl cyclase that elevates the level of intracellular cAMP, to investigate the effect of cAMP and three Rho-family small GTPases (RhoA, Cdc42, and Rac1) on the regulation of cell-cell contact formation in a murine podocyte cell line. Transmission electron microscopy and the immunostaining of cell adhesion molecules and actin-associated proteins have revealed a structural change at the site of cell-cell contact following forskolin treatment. The activity of the Rho-family small GTPases before and after forskolin treatment has been evaluated with a glutathione-S-transferase pull-down assay. Forskolin reinforces the integrity of cell-cell contacts, resulting in the closure of an intercellular adhesion zipper, accompanied by a redistribution of cell adhesion molecules and actin-associated proteins in a continuous linear pattern at cell-cell contacts. The Rho-family small GTPases Rac1 and Cdc42 are activated during closure of the adhesion zipper, whereas RhoA is suppressed. Thus, cAMP promotes the assembly of cell-cell contacts between podocytes via a mechanism that probably involves Rho-family small GTPases.

DOI： 10.1007/s00441-006-0365-3

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The first event occurring at the boundary between a metal implant and living tissue is the attachment of cells onto the metal surface of the implant. The attachment characteristics of the metal in this situation are critical in determining its biocompatibility and usefulness as artificial bone and tooth implants. Using the human osteosarcoma cell line Saos-2, we attempted to establish simple and reliable methods for evaluating the attachment of cultured osteoblastic cells onto titanium samples that had been subjected to various surface treatments. Fluorescence actin imaging showed that cells cultured on titanium with hydrofluoric acid etching (HF-Ti) exhibited delayed spreading of their cytoplasm, as compared to cells cultured for the same length of time on nitrided titanium or physically polished titanium. The HF-Ti-cultured cells also exhibited poor assembly of focal contacts, as visualized by vinculin immunofluorescence. Furthermore, in motility assays based on an in vitro wound model, cells cultured on HF-Ti migrated more slowly than cells cultured on other titanium surfaces. These data suggest that Saos-2 cells attach less effectively to the HF-Ti surface. The methods described in this study should be useful for assessing the initial interactions of cultured cells with various materials, including metals.

DOI： 10.1007/s00441-005-0153-5

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The renal glomerular podocyte exhibits a highly arborized morphology. In comparison with the neuron, which is the best studied process-bearing cell, the podocyte major processes share many cell biological characteristics with neuronal dendrites. Both podocytes and neurons develop microtubule-based thick processes with branching morphology and both have thin actin-based projections (i.e. podocyte foot processes and dendritic spines). Formation of podocyte processes and neuronal dendrites depends on the assembly of microtubules. Because the assembly of microtubules is regulated by phosphorylation of microtubule-associated proteins, inhibition of protein phosphatases abolishes and inhibition of protein kinases promotes process formation. Podocytes and dendrites also share the machinery of intracellular traffic of membranous vesicles, as well as cytoskeletal elements, which is indispensable for the elongation of these processes. Furthermore, these two cell types share expression of various molecules working for signal transduction, transmembranous transport and intercellular contacts. Such common gene expression implies a similar transcriptional regulation in these cells. Concerning the formation of podocyte foot processes and dendritic branches, actin filaments are thought to play a central role in orchestrating the function of various molecules and the regulation of actin assembly is necessary to establish and maintain such sophisticated cellular architecture. The molecular mechanism of foot process formation seems to include Rho family small GTP-binding proteins, which are known to be responsible for the establishment of dendritic branching morphology.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-LISS  

In this review article we discuss the common mechanism for cellular process formation. Besides the podocyte, the mechanism of process formation, including cytoskeletal organization and signal transduction, etc., has been studied using neurons and glias as model systems. There has been an accumulation of data showing common cell biological features of the podocyte and the neuron: 1) Both cells possess long and short cell processes equipped with highly organized cytoskeletal systems; 2) Both show cytoskeletal segregation; microtubules (MTs) and intermediate filaments (IFs) in podocyte primary processes and in neurites, while actin filaments (AFs) are abundant in podocyte foot processes in neuronal synaptic regions; 3) In both cells, process formation is mechanically dependent on MTs, whose assembly is regulated by various microtubule-associated proteins (MAPS); 4) In both cells, process formation is positively regulated by PP2A, a Ser/Thr protein phosphatase; 5) In both cells, process formation is accelerated by laminin, an extracellular matrix protein. In addition, recent data from our and other laboratories have shown that podocyte processes share many features with neuronal dendrites: 1) Podocyte processes and neuronal dendrites possess MTs with mixed polarity, namely, plus-end-distal and minus-end-distal MTs coexist in these processes; 2) To establish the mixed polarity of MTs, both express CHO1/MKLP1, a kinesin-related motor protein, and when its expression is inhibited formation of both podocyte processes and neuronal dendrites is abolished; 3) The elongation of both podocyte processes and neuronal dendrites is supported by rab8-regulated basolateral-type membrane transport; 4) Both podocyte processes and neuronal dendrites express synaptopodin, an actin-associated protein, in a development-dependent manner; interestingly, in both cells, synaptopodin is localized not in the main shaft of processes but in thin short projections from the main shaft. We propose that the podocyte process and the neuronal dendrite share many features, while the neuronal axon should be thought of as an exceptionally differentiated cellular process. (C) 2002 Wiley-Liss, Inc.

DOI： 10.1002/jemt.10077

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The “Model Core Curriculum for Medical Education” (hereinafter referred to as “Core Curriculum”) was revised and released in November 2022. The 2022 Core Curriculum has 10 major items as the basic qualities and abilities required of physicians, and about 600 learning objectives divided into four tiers. The 2022 Core Curriculum should be applied from the students admitted in the 2024 academic year. It is necessary to clarify the correspondence between the currently implemented medical education curriculum and the Core Curriculum. This time, FD training in the form of a workshop was held for faculty members in charge of education at the Ehime University School of Medicine, targeting the pre-clinical curriculum from the first year to the first semester of the fourth year, with the aim of supporting each course in its adaptation to the Core Curriculum. The results showed that while many subjects were applicable to several areas, relatively few classes were applicable to one of the major items “Medicine in Society”. Furthermore, there were no applicable classes for 3 items at the fourth tire of the Core Curriculum. We suggest that the Department of Medicine systematically examine this curriculum improvement.

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Prosaposin (PSAP), a highly conserved glycoprotein, is a precursor of saposins A-D. Accumulating evidence suggests that PSAP is a neurotrophic factor, as well as a regulator of lysosomal enzymes. Recently, the orphan G-protein-coupled receptors GPR37 and GPR37L1 were recognized as PSAP receptors, but their functions have not yet been clarified. In this study, we examined the distribution of PSAP and its receptors in the dorsal root ganglion (DRG) during development using specific antibodies, and showed that PSAP accumulates primarily in lysosomes and is dispersed throughout the cytoplasm of satellite cells. Later, PSAP colocalized with two receptors in satellite cells, and formed a characteristic ring shape approximately 8 weeks after birth, during a period of rapid DRG development. This ring shape, which was only observed around larger neurons, is evidence that several satellite cells are synchronously activated. We found that sortilin, a transporter of a wide variety of intracellular proteins containing PSAP, is strongly localized to the inner side of satellite cells, which contact the neuronal surface. These findings suggest that PSAP and GPR37/GPR37L1 play a role in activating both satellite and nerve cells.

DOI： 10.1371/journal.pone.0255958

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Authorship：Last author   Publishing type：Research paper (bulletin of university, research institution)   Publisher：愛媛大学教育・学生支援機構  

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Prosaposin (PSAP) has two forms: a precursor and a secreted form. The secreted form has neurotrophic, myelinotrophic, and myotrophic properties. The precursor form is a precursor protein of saposins A-D. Although the distribution of PSAP in male reproductive organs is well known, its distribution in female reproductive organs, especially in the oviduct, is unclear. Immunoblots and immunohistochemistry of oviducts showed that oviductal tissues contain PSAP proteins, and a significant increase in PSAP was observed in the estrus-metestrus phase compared to the diestrus-proestrus phase in the ampulla. To identify PSAP trafficking in cells, double-immunostaining was performed with antibodies against PSAP in combination with sortilin, mannose 6 phosphate receptor (M6PR), or low-density lipoprotein receptor-related protein 1 (LRP1). PSAP and sortilin double-positive reactions were observed near the nuclei, as well as in the apical portion of microvillous epithelial cells, whereas these reactions were only observed near the nuclei of ciliated epithelial cells. PSAP and M6PR double-positive reactions were observed near the nuclei of microvillous and ciliated epithelial cells. PSAP and M6PR double-positive reactions were also observed in the apical portion of microvillous epithelial cells. PSAP and LRP1 double-positive reactions were observed in the plasma membrane and apical portion of both microvillous and ciliated epithelial cells. Immunoelectron staining revealed PSAP immunoreactive small vesicles with exocytotic features at the apical portion of microvillous epithelial cells. These findings suggest that PSAP is present in the oviductal epithelium and has a pivotal role during pregnancy in providing an optimal environment for gametes and/or sperm in the ampulla.

DOI： 10.1007/s00441-020-03339-x

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Authorship：Last author   Language：Japanese   Publishing type：Research paper (bulletin of university, research institution)  

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Authorship：Last author   Publishing type：Research paper (bulletin of university, research institution)   Publisher：愛媛大学教育・学生支援機構  

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Salivary glands produce various neurotrophins that are thought to regulate salivary function during normal and pathological conditions. Prosaposin (PSAP) is a potent neurotrophin found in several tissues and various biological fluids and may play roles in the regulation of salivary function. However, little is known about PSAP in salivary glands. As the functions of salivary glands are diverse based on age and sex, this study examines whether PSAP and its receptors, G protein-coupled receptor 37 (GPR37) and GPR37L1, are expressed in the salivary glands of rats and whether sex and aging affect their expression. Immunohistochemical analysis revealed that PSAP and its receptors were expressed in the major salivary glands of rats, although their expression varied considerably based on the type of gland, acinar cells, age and sex. In fact, PSAP, GPR37 and GPR37L1 were predominantly expressed in granular convoluted tubule cells of the submandibular gland and the intensity of their immunoreactivity was higher in young adult female rats than age-matched male rats, which was more prominent at older ages (mature adult to menopause). On the other hand, weak PSAP, GPR37 and GPR37L1 immunoreactivity was observed mainly in the basal layer of mucous cells of the sublingual gland. Triple label immunofluorescence analysis revealed that PSAP, GPR37 and GPR37L1 were co-localized in the basal layer of acinar and ductal cells in the major salivary glands. The present findings indicate that PSAP and its receptors, GPR37 and GPR37L1, are expressed in the major salivary glands of rats and their immunoreactivities differ considerably with age and sex.

DOI： 10.1007/s00441-018-2835-9

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Prosaposin (PS) is a secretory neurotrophic factor, as well as a regulator of lysosomal enzymes. We previously reported the up-regulation of PS and the possibility of its axonal transport by GABAergic interneurons after exocitotoxicity induced by kainic acid (KA), a glutamate analog. In the present study, we performed double immunostaining with PS and three calcium binding protein markers: parvalbumin (PV), calbindin, and calretinin, for the subpopulation of GABAergic interneurons, and clarified that the increased PS around the hippocampal pyramidal neurons after KA injection existed mainly in the axons of PV positive interneurons. Electron microscopy revealed PS containing vesicles in the PV positive axon. Double immunostaining with PS and secretogranin or synapsin suggested that PS is secreted with secretogranin from synapses. Based on the results from in situ hybridization with two alternative splicing forms of PS mRNA, the increase of PS in the interneurons was due to the increase of PS + 0 (mRNA without 9-base insertion) as in the choroid plexus, but not PS + 9 (mRNA with 9-base insertion). These results were similar to those from the choroid plexus, which secretes an intact form PS + 0 to the cerebrospinal fluid. Neurons, especially PV positive GABAergic interneurons, produce and secrete the intact form of PS around hippocampal pyramidal neurons to protect them against KA neurotoxicity.

DOI： 10.1016/j.ibror.2017.07.001

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Language：Japanese   Publisher：Japan Society for Pharmaceutical Education  

<p>The physical assessment of vital signs and related parameters has recently been introduced into the curriculum of many pharmacy schools throughout Japan. To utilize the acquired skills in actual clinical settings, an accurate clinical assessment of a patient's condition is essential. In the present study, pharmacy and medical students completed for the first time a Basic Physiology of Vital Signs (BPVS) program, which is a basic-level course in the Clinical Physiology of Vital Signs program developed for medical residency, and the educational outcomes among students were evaluated. Both groups of students acquired in repeated trials the ability to perform collection, handling, and evaluation of clinical information in a limited time while considering various clinical priorities based on their developing knowledge. This finding suggests that the BPVS program is an effective learning method for pharmacy and medical students that facilitates clinical diagnosis.</p>

DOI： 10.24489/jjphe.2017-005

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<p>Introduction: Compared with faculties in clinical and medical research departments, those in medical departments are not appropriately evaluated in terms of their contributions to or achievements in medical education. Therefore, the aims of this study were to investigate the contributions of medical department faculties to medical education, and to examine differences in contributions according to duty positions and specialties.</p><p>Methods: Five-grade self-assessments in relation to 20 items on a rating form for performance in medical education, which was developed by the Japan Society for Medical Education's Committee for Performance Evaluation, were carried out by medical department faculties in Japanese universities. The data were then totalized and analyzed.</p><p>Results and Discussion: Although faculties belonging to departments other than medical education units did not actively participate in examinations or the education system, they still made contributions to lectures and practice. In addition, faculties with positions with more duties tended to show greater participation in the education system.</p><p></p><p>Conclusion: Based on these findings, we recommend the use of a rating form as a standard scale to evaluate performance in medical education.</p>

DOI： 10.11307/mededjapan.47.2_77

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Other Link： http://search.jamas.or.jp/link/ui/2016270200

Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

Four sphingolipid activator proteins (i.e., saposins A-D) are synthesized from a single precursor protein, prosaposin (PS), which exerts exogenous neurotrophic effects in vivo and in vitro. Kainic acid (KA) injection in rodents is a good model in which to study neurotrophic factor elevation; PS and its mRNA are increased in neurons and the choroid plexus in this animal model. An 18-mer peptide (LSELIINNATEELLIKGL; PS18) derived from the PS neurotrophic region prevents neuronal damage after ischemia, and PS18 is a potent candidate molecule for use in alleviating ischemia-induced learning disabilities and neuronal loss. KA is a glutamate analog that stimulates excitatory neurotransmitter release and induces ischemia-like neuronal degeneration; it has been used to define mechanisms involved in neurodegeneration and neuroprotection. In the present study, we demonstrate that a subcutaneous injection of 0.2 and 2.0 mg/kg PS18 significantly improved behavioral deficits of Wistar rats (n = 6 per group), and enhanced the survival of hippocampal and cortical neurons against neurotoxicity induced by 12 mg/kg KA compared with control animals. PS18 significantly protected hippocampal synapses against KA-induced destruction. To evaluate the extent of PS18- and KA-induced effects in these hippocampal regions, we performed histological evaluations using semithin sections stained with toluidine blue, as well as ordinal sections stained with hematoxylin and eosin. We revealed a distinctive feature of KA-induced brain injury, which reportedly mimics ischemia, but affects a much wider area than ischemia-induced injury: KA induced neuronal degeneration not only in the CA1 region, where neurons degenerate following ischemia, but also in the CA2, CA3, and CA4 hippocampal regions.

DOI： 10.1371/journal.pone.0126856

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Four sphingolipid activator proteins (i.e., saposins A-D) are synthesized from a single precursor protein, prosaposin (PS), which exerts exogenous neurotrophic effects in vivo and in vitro. Kainic acid (KA) injection in rodents is a good model in which to study neurotrophic factor elevation; PS and its mRNA are increased in neurons and the choroid plexus in this animal model. An 18-mer peptide (LSELIINNATEELLIKGL; PS18) derived from the PS neurotrophic region prevents neuronal damage after ischemia, and PS18 is a potent candidate molecule for use in alleviating ischemia-induced learning disabilities and neuronal loss. KA is a glutamate analog that stimulates excitatory neurotransmitter release and induces ischemia-like neuronal degeneration; it has been used to define mechanisms involved in neurodegeneration and neuroprotection. In the present study, we demonstrate that a subcutaneous injection of 0.2 and 2.0 mg/kg PS18 significantly improved behavioral deficits of Wistar rats (n = 6 per group), and enhanced the survival of hippocampal and cortical neurons against neurotoxicity induced by 12 mg/kg KA compared with control animals. PS18 significantly protected hippocampal synapses a

DOI： 10.1371/journal.pone.0126856

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Because excessive glutamate release is believed to play a pivotal role in numerous neuropathological disorders, such as ischemia or seizure, we aimed to investigate whether intrinsic prosaposin (PS), a neuroprotective factor when supplied exogenously in vivo or in vitro, is up-regulated after the excitotoxicity induced by kainic acid (KA), a glutamate analog. In the present study, PS immunoreactivity and its mRNA expression in the hippocampal and cortical neurons showed significant increases on day 3 after KA injection, and high PS levels were maintained even after 3 weeks. The increase in PS, but not saposins, detected by immunoblot analysis suggests that the increase in PS-like immunoreactivity after KA injection was not due to an increase in saposins as lysosomal enzymes after neuronal damage, but rather to an increase in PS as a neurotrophic factor to improve neuronal survival. Furthermore, several neurons with slender nuclei inside/outside of the pyramidal layer showed more intense PS mRNA expression than other pyramidal neurons. Based on the results from double immunostaining using anti-PS and anti-GABA antibodies, these neurons were shown to be GABAergic interneurons in the extraand intra-pyramidal layers. In the cerebral cortex, several large neurons in the V layer showed very intense PS mRNA expression 3 days after KA injection. The choroid plexus showed intense PS mRNA expression even in the normal rat, and the intensity increased significantly after KA injection. The present study indicates that inhibitory interneurons as well as stimulated hippocampal pyramidal and cortical neurons synthesize PS for neuronal survival, and the choroid plexus is highly activated to synthesize PS, which may prevent neurons from excitotoxic neuronal damage. To the best of our knowledge, this is the first study that demonstrates axonal transport and increased production of neurotrophic factor PS after KA injection.

DOI： 10.1371/journal.pone.0110534

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Neurogenesis in the hippocampal dentate gyrus occurs constitutively throughout postnatal life. Adult neurogenesis includes a multistep process that ends with the formation of a postmitotic and functionally integrated new neuron. During adult neurogenesis, various markers are expressed, including GFAP, nestin, Pax6, polysialic acid-neural cell adhesion molecule (PSA-NCAM), neuronal nuclei (NeuN), doublecortin, TUC-4, Tuj-1, and calretinin. Prosaposin is the precursor of saposins A-D; it is found in various organs and can be excreted. Strong prosaposin expression has been demonstrated in the developing brain including the hippocampus, and its neurotrophic activity has been proposed. This study investigated changes in prosaposin in the dentate gyrus of young and adult rats using double immunohistochemistry with antibodies to prosaposin, PSA-NCAM, and NeuN. Prosaposin immunoreactivity was intense in the dentate gyrus at postnatal day 3 (P3) and P7, but decreased gradually after P14. In the dentate gyrus at P28, immature PSA-NCAM-positive neurons localized exclusively in the subgranular zone were prosaposin-negative, whereas mature Neu-N-positive neurons were positive for prosaposin. Furthermore, these prosaposin-negative immature neurons were saposin B-positive, suggesting that the neurons take up and degrade prosaposin. In situ hybridization assays showed that prosaposin in the adult dentate gyrus is dominantly the Pro+9 type, a secreted type of prosaposin. These results imply that prosaposin secreted from mature neurons stimulates proliferation and maturation of immature neurons in the dentate gyrus.

DOI： 10.1371/journal.pone.0095883

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Prosaposin has two distinct profiles. One is a precursor form that is processed into saposins thus promoting lysosomal sphingolipid hydrolase function, whereas the other is an intact form that is not processed into saposins but isabundant in certain tissues and secretory fluids, including the cerebrospinal fluid. In rats, alternative splicing in the prosaposin gene generates mRNAs with and without a 9-base insertion (Pro+9 and Pro+0 mRNAs, respectively). Pro+9 mRNA is reported to be preferentially expressed in tissues in which the intact form of prosaposin dominates, whereas Pro+0 mRNA is preferentially expressed in tissues in which the precursor dominates. The expression patterns of Pro+9 and Pro+0 mRNAs in the rat choroid plexus are examined in the present study. The specificities of 36-mer oligonucleotide probes used to detect the 9-base insertion by in situ hybridization were demonstrated by dot-blot hybridization. Next, these probes were used for in situ hybridization, which showed predominant expressionof Pro+0 mRNA and weak expression of Pro+9 mRNA in the choroid plexus. These expression patterns were confirmed by reverse transcription plus the polymerase chain reaction with

DOI： 10.1007/s00441-013-1773-9

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

Prosaposin has two distinct profiles. One is a precursor form that is processed into saposins thus promoting lysosomal sphingolipid hydrolase function, whereas the other is an intact form that is not processed into saposins but is abundant in certain tissues and secretory fluids, including the cerebrospinal fluid. In rats, alternative splicing in the prosaposin gene generates mRNAs with and without a 9-base insertion (Pro+9 and Pro+0 mRNAs, respectively). Pro+9 mRNA is reported to be preferentially expressed in tissues in which the intact form of prosaposin dominates, whereas Pro+0 mRNA is preferentially expressed in tissues in which the precursor dominates. The expression patterns of Pro+9 and Pro+0 mRNAs in the rat choroid plexus are examined in the present study. The specificities of 36-mer oligonucleotide probes used to detect the 9-base insertion by in situ hybridization were demonstrated by dot-blot hybridization. Next, these probes were used for in situ hybridization, which showed predominant expression of Pro+0 mRNA and weak expression of Pro+9 mRNA in the choroid plexus. These expression patterns were confirmed by reverse transcription plus the polymerase chain reaction with AlwI restriction enzyme treatment. Expression of the intact form of prosaposin in the choroid plexus was assessed by Western blotting and immunohistochemistry. Because the choroid plexus is responsible for the generation of cerebrospinal fluid containing the intact form of prosaposin, the present study raises the possibility that Pro+0 mRNA is related to the intact form in the choroid plexus and that the alternatively spliced forms of mRNAs do not simply correspond to the precursor and intact forms of prosaposin.

DOI： 10.1007/s00441-013-1773-9

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Language：Japanese   Publisher：(一社)日本医学教育学会  

目的:医学研究科の大学院生が希望する研究指導の内容について男女差を評価した。方法:2011年12月から2012年1月の間に、6大学院医学研究科で大学院生1,700人を対象に質問紙調査を行った。研究教育における複数の指導項目や指導体制を提示し、各項目の希望者の割合を男女で比較した。結果:回答者は676人(女性227人)であった。女性は男性よりキャリア形成、コンピュータ、統計解析の指導を希望する者が多かった。また、男性は女性よりも指導者との関係は上下関係(師弟関係)が望ましいと回答した人が多かった。結論:女性医師の増加に伴い、性別によるニーズの違いに配慮した研究教育の検討が望まれる。(著者抄録)

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Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2014&ichushi_jid=J00050&link_issn=&doc_id=20140310010001&doc_link_id=%2Fed9jmded%2F2014%2F004501%2F001%2F0001-0007%26dl%3D0&url=http%3A%2F%2Fwww.medicalonline.jp%2Fjamas.php%3FGoodsID%3D%2Fed9jmded%2F2014%2F004501%2F001%2F0001-0007%26dl%3D0&type=MedicalOnline&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00004_2.gif

Language：English   Publishing type：Research paper (scientific journal)   Publisher：BIOMED CENTRAL LTD  

Because WT1 is expressed in leukemia cells, the development of cancer immunotherapy targeting WT1 has been an attractive translational research topic. However, concern of this therapy still remains, since WT1 is abundantly expressed in renal glomerular podocytes. In the present study, we clearly showed that WT1-specific cytotoxic T lymphocytes (CTLs) certainly exerted cytotoxicity against podocytes in vitro; however, they did not damage podocytes in vivo. This might be due to the anatomical localization of podocytes, being structurally separated from circulating CTLs in glomerular capillaries by an exceptionally thick basement membrane.

DOI： 10.1186/1756-8722-7-3

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：PUBLIC LIBRARY SCIENCE  

Background: Duchenne muscular dystrophy caused by a mutation in the X-linked dystrophin gene induces metabolic and structural disorders in the brain. A lack of dystrophin in brain structures is involved in impaired cognitive function. Prosaposin (PS), a neurotrophic factor, is abundant in the choroid plexus and various brain regions. We investigated whether PS serves as a link between dystrophin loss and gross and/or ultrastructural brain abnormalities.
Methodology/Principal Findings: The distribution of PS in the brains of juvenile and adult mdx mice was investigated by immunochemistry, Western blotting, and in situ hybridization. Immunochemistry revealed lower levels of PS in the cytoplasm of neurons of the cerebral cortex, hippocampus, cerebellum, and choroid plexus in mdx mice. Western blotting confirmed that PS levels were lower in these brain regions in both juveniles and adults. Even with low PS production in the choroids plexus, there was no significant PS decrease in cerebrospinal fluid (CSF). In situ hybridization revealed that the primary form of PS mRNA in both normal and mdx mice was Pro+ 9, a secretory-type PS, and the hybridization signals for Pro+ 9 in the above-mentioned brain regions were weaker in mdx mice than in normal mice. We also investigated mitogen-activated protein kinase signalling. Stronger activation of ERK1/2 was observed in mdx mice, ERK1/2 activity was positively correlated with PS activity, and exogenous PS18 stimulated both p-ERK1/2 and PS in SH-SY5Y cells.
Conclusions/Significance: Low levels of PS and its receptors suggest the participation of PS in some pathological changes in the brains of mdx mice.

DOI： 10.1371/journal.pone.0080032

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：F HERNANDEZ  

The trophic factor prosaposin (PS) is strongly expressed in skeletal muscle, and reportedly, a PS-derived peptide attenuates loss of muscle mass after nerve injury in vivo and increases myoblast fusion into myotubes in vitro. However, few studies have focused on the role of PS during muscle regeneration. We examined the expression of PS in the skeletal muscles in normal, mdx, and cardiotoxin (CTX)-treated mice using immunofluorescence staining, Western blotting, and in situ hybridisation. Immunofluorescence showed intense PS immunoreactivity in the peripheral cytoplasm of uninjured myofibres of normal mice and regenerated myofibres of 8 weeks post-CTX-injection mice. In early stage CTX-treated mice (14 days and earlier), intense PS immunoreactivity was also detected in the immune cells that infiltrated damaged muscle, but it was weak for regenerating myofibres. Western blot confirmed these findings. In contrast, PS was continuously low in mdx mice in both immunofluorescence and Western blotting. In situ hybridisation confirmed the decrease of PS mRNA in regenerated myofibres and revealed the main form of PS mRNA as Pro+0 without a 9-base insertion both in normal and mdx mice. The embryonic myosin (MYH3) was clearly localized in the newly regenerated myofibres at 3, 7, and 14 days of post-CTX-injection and mdx mice, but was lower in the late stage of regenerated myofibres (28 and 56 days post-CTX injection). The inverse distribution of MYH3 and PS indicates that the PS expression is closely related to the differentiation of regenerated myofibres. Investigation of the mitogen-activated protein (MAP) kinase signal pathway showed the inversely synchronous correlation of phosphorylated ERK1/2 with myofibre PS and the synchronous correlation of phosphorylated p-38 with myofibre PS. These data suggest that PS is involved in the regulation of muscle differentiation of regenerated fibres.

DOI： 10.14670/HH-28.875

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Language：English   Publishing type：Research paper (scientific journal)  

Prosaposin (PSAP) is as a trophic factor and an activator protein for sphingolipid hydrolase in lysosomes. We generated a specific antibody to PSAP and examined the spatiotemporal distribution of PSAP-immunoreactive (PSAP-IR) cells in the lymphatic tissues of Wistar rats. Immunoblots of tissue homogenates separated electrophoretically showed a single band for PSAP in brain but two bands in spleen. PSAP-IR cells were distributed in both the red and white pulp of the spleen, in both the cortex and medulla of the thymus and in mesenteric lymph nodes. Many PSAP-IR cells were found in the dome portion of Peyer's patches and the number of PSAP-IR cells increased with the age of the rat. To identify the PSAP-IR cells, double- and triple-immunostainings were performed with antibodies against PSAP, CD68 and CD1d. The large number of double- and triple-positive cells suggested that antigen-presenting cells contained much PSAP in these lymphatic tissues. Intense expression of PSAP mRNA, examined by in situ hybridisation, was observed in the red pulp and corona of the spleen. In rats, the PSAP gene generates two alternative splicing forms of mRNA: Pro+9 containing a 9-base insertion and Pro+0 without the insertion. We examined the expression patterns of the alternative splicing forms of PSAP mRNA in the spleen. The presence of both types of mRNA (Pro+9 and Pro+0) indicated that the spleen contains various types of prosaposin-producing and/or secreting cells. These findings suggest diverse functions for PSAP in the immune system. © 2013 Springer-Verlag Berlin Heidelberg.

DOI： 10.1007/s00441-013-1575-0

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Language：English   Publishing type：Research paper (scientific journal)  

An 18-merpeptide derived from the neurotrophic region of prosaposin (PS-pep) prevents hearing loss and cochlear damage due to transient cochlear ischaemia by activating an anti-apoptotic pathway. PS-pep is a potent candidate molecule for alleviating ischaemia-induced hearing loss.PS-pep was investigated for its protective effects against ischaemia-induced hearing loss and cochlear damage.Ischaemia was induced in both cochleae in Mongolian gerbils by pulling the ligatures around both vertebral arteries in an anterior direction using 5 g weights for 15 min. PS-pep was synthesized artificially and administered subcutaneously four times after the induction of transient cochlear ischaemia.An increase in the auditory brainstem response threshold was alleviated in animals treated with 2.0 mg/kg PS-pep. Histological examinations conducted on day 7 showed that the loss of inner hair cells (IHCs) was more prominent than that of outer hair cells. Higher doses of PS-pep significantly alleviated IHC loss. An increase in the anti-apoptotic factor bcl-2 was also noted in the IHCs treated with PS-pep.

DOI： 10.3109/00016489.2012.750430

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：INFORMA HEALTHCARE  

Conclusion: An 18-mer peptide derived from the neurotrophic region of prosaposin (PS-pep) prevents hearing loss and cochlear damage due to transient cochlear ischaemia by activating an anti-apoptotic pathway. PS-pep is a potent candidate molecule for alleviating ischaemia-induced hearing loss. Objective: PS-pep was investigated for its protective effects against ischaemia-induced hearing loss and cochlear damage. Methods: Ischaemia was induced in both cochleae in Mongolian gerbils by pulling the ligatures around both vertebral arteries in an anterior direction using 5 g weights for 15 min. PS-pep was synthesized artificially and administered subcutaneously four times after the induction of transient cochlear ischaemia. Results: An increase in the auditory brainstem response threshold was alleviated in animals treated with 2.0 mg/kg PS-pep. Histological examinations conducted on day 7 showed that the loss of inner hair cells (IHCs) was more prominent than that of outer hair cells. Higher doses of PS-pep significantly alleviated IHC loss. An increase in the anti-apoptotic factor bcl-2 was also noted in the IHCs treated with PS-pep.

DOI： 10.3109/00016489.2012.750430

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Language：English   Publishing type：Research paper (scientific journal)  

Duchenne muscular dystrophy caused by a mutation in the X-linked dystrophin gene induces metabolic and structural disorders in the brain. A lack of dystrophin in brain structures is involved in impaired cognitive function. Prosaposin (PS), a neurotrophic factor, is abundantin the choroid plexus and various brain regions. We investigated whether PS serves as a link between dystrophin loss and gross and/or ultrastructural brain abnormalities.The distribution of PS in the brains of juvenile and adult mdx mice was investigated by immunochemistry, Western blotting, and in situ hybridization. Immunochemistry revealed lower levels of PS in the cytoplasm of neurons of the cerebral cortex, hippocampus, cerebellum, and choroid plexus in mdx mice. Western blottingconfirmed that PS levels were lower in these brain regions in both juveniles and adults. Even with low PS production in the choroids plexus,there was no significantPS decrease in cerebrospinal fluid (CSF). Insitu hybridization revealed that the primary form of PS mRNA in both normaland mdx mice was Pro+9, a secretory-typePS, and the hybridization signals for Pro+9 in the above-mentioned brain regions were weaker in mdx mice than in n

DOI： 10.1371/journal.pone.0080032

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Language：Japanese  

CiNii Books

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Other Link： https://opac1.lib.ehime-u.ac.jp/iyokan/TD00001083

Language：Japanese  

CiNii Books

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Other Link： http://iyokan.lib.ehime-u.ac.jp/dspace/handle/iyokan/1595

Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC VET SCI  

The stomach of the Pacific white-sided dolphin is divided into three parts: forestomach, proper gastric gland portion, and pyloric chamber. The histological features of the dolphin stomach are similar to those of terrestrial mammal stomachs, although the distribution of glycoconjugates in mucosal cells of the dolphin stomach is unknown. To learn about glycoconjugates in cetacean gastric mucosa, the glycoconjugate distribution in the mucous epithelium of the Pacific white-sided dolphin was studied using 21 lectins. Among the lectins tested, GSL-I and DBA specifically labelled the superficial layer of the forestomach epithelium. GSL-I, SBA, RCA-I, VVA, GSL-I I, DSL, LEL, STL, s-WGA, WGA. PNA, and Jacalin labelled the luminal surface of the chief cells in the proper gastric gland. GSL-I, SBA, RCA-I, DSL, LEL, STL, s-WGA, PNA, and LCA labelled tubular structures in the cytoplasm of parietal cells. The surface portion of the pits in the pyloric chamber strongly reacted with RCA-I, GSL-II, WGA, PNA, LCA, PHA-L, and UEA-I, whereas the neck portion reacted weakly. Although lining one tubular portion, individual secretory cells in the pyloric gland displayed a heterogeneous reaction. This is the first report on the lectin histochemistry of a cetacean stomach and reveals GSL-I and DBA as specific marker lectins for the cornified stratified squamous epithelium cells of the Pacific white-sided dolphin. The stomachs of cetaceans and terrestrial mammals have similar histological features and mucous glycoconjugate content.

DOI： 10.1292/jvms.11-0115

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Language：Japanese  

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：愛媛医学会  

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Prosaposin is the precursorprotein of four glycoproteins, saposins A, B, C, andD, which activate sphingolipid hydrolases in lysosomes. Besides this role, intact prosaposin is also known as a potent neurotrophic factor that prevents neuronal cell death and stimulates neurite outgrowth in in vivo and in vitro experiments. In the present study, we examined chronological changes in prosaposin immunoreactivity in the rat brain using immunofluorescence staining and Diaminobenzidine (DAB) immunohistochemistry. In the hippocampal regions CA1, CA3, and dentate gyrus, the strongest stainingof prosaposin was observed on postnatal day 1. The prosaposin immunoreactivity then decreased gradually until postnatal day 28. But in the cerebral cortex, prosaposin staining intensity increased from postnatal day 1 to 14, then decreased until postnatal day 28. The prosaposin immunoreactivity co-localized with the lysosomal granules labeled by an anti-Cathepsin D antibody, indicating that prosaposin mainly localized in the lysosomes of the neurons. We alsoexamined the chronological changes in prosaposin mRNA and its two alternatively spliced variants using in situ hybridization. We found that both the mRN

DOI： 10.1016/j.neures.2011.06.001

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER IRELAND LTD  

Spina bifida aperta (SBA) is an open neural tube defect that occurs during the embryonic period. We created SBA chicks by incising the roof plate of the neural tube in the embryo. The area of the dorsal funiculus was smaller in the SBA chicks than in the normal controls. Additionally, the SBA group had fewer nerve fibres in the dorsal funiculus than the normal controls. The pathway of the ascending sensory nerves was revealed by tracing the degenerated nerve fibres using osmification. We cut the sciatic nerve (L5) of the control and SBA chicks at the central end of the dorsal root ganglion 1 day after hatching and fixed the tissue 3 days later. Degenerated sensory nerve fibres were observed in the ipsilateral dorsal funiculus in the control chicks. In contrast, degenerated sensory nerve fibres were observed in the ipsilateral and contralateral dorsal, ventral and lateral funiculi of the spinal cord in the SBA chicks. Consequently, fewer sensory nerve fibres ascended to the thoracic dorsal funiculus in the SBA chicks than in the normal controls. This is the first report of abnormal changes in the ascending sensory nerve fibres in SBA. (C) 2011 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

DOI： 10.1016/j.neures.2011.05.017

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Prosaposin is the precursor protein of four glycoproteins, saposins A, B, C, and D, which activate sphingolipid hydrolases in lysosomes. Besides this role, intact prosaposin is also known as a potent neurotrophic factor that prevents neuronal cell death and stimulates neurite outgrowth in in vivo and in vitro experiments. In the present study, we examined chronological changes in prosaposin immunoreactivity in the rat brain using immunofluorescence staining and Diaminobenzidine (DAB) immunohistochemistry. In the hippocampal regions CA1, CA3, and dentate gyrus, the strongest staining of prosaposin was observed on postnatal day 1. The prosaposin immunoreactivity then decreased gradually until postnatal day 28. But in the cerebral cortex, prosaposin staining intensity increased from postnatal day 1 to 14, then decreased until postnatal day 28. The prosaposin immunoreactivity co-localized with the lysosomal granules labeled by an anti-Cathepsin D antibody, indicating that prosaposin mainly localized in the lysosomes of the neurons. We also examined the chronological changes in prosaposin mRNA and its two alternatively spliced variants using in situ hybridization. We found that both the mRNA forms, especially the one without a nine-base insertion, increased significantly from embryonic day 15 to postnatal day 7, then decreased gradually until postnatal day 28. Abundant prosaposin expression in the perinatal stages indicates a potential role of prosaposin in the early development of the rat brain. © 2011 Elsevier Ireland Ltd and the Japan Neuroscience Society.

DOI： 10.1016/j.neures.2011.06.001

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC VET SCI  

Sugars in the glycocalyx play an important role in the attachment of infectious agents to the respiratory mucosa. We examined the histochemistry of 23 lectins to survey the sugar expression in the glycocalyx of the respiratory mucosa of the Pacific white-sided dolphin. Lagenorhynchus obliquidens. The ciliated and basal cells were positive for all of the lectins studied. SBA, WFA, GSL-II, STL, S-WGA, and PNA staining in the cytoplasm showed different intensities between basal cells and ciliated cells. These results suggest that multiple terminal glycosylation occurs on ciliated and basal cells, such as GaINAc, GIcNAc, NeuNAc, galactose, glucose/mannose, olieosaccharide, and fucose, and that sugar residue expression changes during cell differentiation. The Pacific white-sided dolphin respiratory mucosa might express multiple sugar residues in the glycocalyx, to prevent the attachment and colonisation of infectious agents.

DOI： 10.1292/jvms.11-0116

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPAN SOC VET SCI  

Spina bifida aperta (SBA) is a congenital malformation of the spinal cord with complications such as spinal ataxia and bowel and bladder dysfunction. We have developed a chick model with surgery-induced SBA that shows spinal ataxia after hatching. In the present study, motor neurons in the early stages in chicks with and without SBA were observed by immunohistochemical staining with a monoclonal antibody against Islet-1, a motor neuron marker. Delay in migration and maturation of motor neurons was observed in SBA. Although the final numbers of Islet-1-positive neurons in these two groups were not different, a detect in the production and elimination of excess motor neurons in the early developmental stages in the SBA group may be involved in the pathological mechanism of the motor complications of this disease.

DOI： 10.1292/jvms.10-0385

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DOI： 10.1292/jvms.10-0385

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DOI： 10.2147/AMEP.S22948

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MARINE BIOLOGICAL LABORATORY  

In echinoderms, the circumesophageal muscle is mesodermal in origin. Several studies of sea urchins have reported that the molecular events of myogenesis occur during the differentiation of the circumesophageal muscle in early embiyogenesis. In contrast, few detailed reports have examined the differentiation of the circumesophagus muscle in larval starfish. Here, we examined the temporal-numeric distribution and differentiation of esophagus circular muscle fibers in the starfish Patina pectinifera by using rhodamine phalloidin staining. Muscle fibers were not detected in mouth-forming larvae, but a mean of about 10 muscle fibers was observed in 48-h larvae, and about 26 bundles were observed after 60 h. During the next 12 h, the number of muscle fiber bundles increased slightly to about 31 bundles and was stable until 96 h.

DOI： 10.1086/BBLv219n1p12

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：OXFORD UNIV PRESS  

Background. Childhood-onset systemic lupus erythematosus (SLE) is frequently complicated with lupus nephritis (LN), which is characterized by the deposition of DNA-containing immune complex to the glomerulus. Toll-like receptor 9 (TLR9), capable of recognizing the microbially derived CpG oligonucleotide, plays a crucial role in the innate immunity. TLR9 is also assumed to be related to the aetiology of SLE in the recognition of anti-DNA antibody-containing immune complex, but this remains controversial. We conducted a study to elucidate the association between TLR9 and LN in childhood-onset SLE.
Methods. We compared the expression and localization of TLR9 and the slit membrane-related protein in the biopsied kidney sample by immunostaining in four children with active or inactive LN. We also evaluated their laboratory findings, such as anti-DNA antibody, complement and proteinuria at biopsy, to assess the correlation to the findings of the immunostaining. Results. TLR9 is not expressed in a normal control kidney. However, TLR9 develops in podocytes only in active LN but disappears in remission. Meanwhile, the slit membrane-related proteins such as nephrin, podocin and synaptopodin in podocytes express clearly and uniformly in remission, but their expression is markedly diminished in active LN, which results in podocyte injury. When TLR9 is expressed in podocytes, all the patients simultaneously showed hypocomplementaemia, high titre of anti-double-stranded DNA (dsDNA) antibody and proteinuria.
Conclusion. Injured podocytes in active LN express TLR9. This expression could be associated with proteinuria and increased anti-dsDNA antibody. This is the first report indicating that TLR9 is involved in the aetiology of LN and that it may play some role in podocyte injury.

DOI： 10.1093/ndt/gfq058

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER  

To examine embryogenic mechanisms in the starfish Patiria (Asterina) pectinifera, we histochemically analyzed several larval stages using Alcian Blue (AB, which stains acidic mucins), Periodic Acid Schiff (PAS, which stains neutral mucins), and 21 types of lectins. Carbohydrate distribution patterns were observed in the cytoplasm, basement membrane, and blastocoel as follows: (1) The first group of lectins showed granular signals in the mesendodermal cells, and these lectins may be useful as mesendoderm markers. (2) The second class of lectins showed diffuse signals across the entire cytoplasm from the hatched blastula until the mid gastrula. These signals became localized to the basal cytoplasm of archenteron cells at the early bipinnaria. (3) Lectin reactivity in the basement membrane peaked at the early-to-mid gastrula and was nearly gone by the early bipinnaria. These results suggest the existence of various substances in the basement membrane and imply the importance of these substances during archenteron elongation and the induction of mesenchyme differentiation. (4) Signal colors with AB-PAS double staining in the blastocoel changed from magenta (by PAS staining) into blue (by AB staining) during these stages, thus, indicating that mucin located in the blastocoel changed from neutral to acidic. The most significant part of this report is the first description regarding temporal changes in the characteristics of intra- and extracellular components with the combination of many different lectins and stains.

DOI： 10.1007/s00427-009-0280-1

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

Recurrent focal segmental glomerulosclerosis (FSGS) is a major challenge in the field of transplantation. Integrin-linked kinase (ILK) has emerged as a key mediator of podocyte-glomerular basement membrane (GBM) interactions. To clarify the involvement of plasma factors in FSGS recurrence, we examined the effects of plasma from FSGS patients with or without posttransplant recurrence on cultured podocytes, focusing particularly on ILK activity. Podocytes from a conditionally immortalized mouse podocyte cell line were treated with plasma from 11 FSGS patients, and ILK activity was determined using an immune complex kinase assay. Treatment with plasma from three patients with recurrence induced an increase in ILK activity. In contrast, no increase in ILK activity was observed in cultured podocytes treated with plasma from the remaining three patients with recurrence and five patients without recurrence. Cultured podocytes treated with plasma that induced ILK activity showed alterations of focal contact and detachment from the laminin matrix. In conclusion, this preliminary study provides experimental evidence suggesting the possible presence of circulating toxic factors in the plasma of some patients with recurrent FSGS, which induce an increase in podocyte ILK activity that may lead to the detachment of podocytes from the GBM.

DOI： 10.1111/j.1600-6143.2008.02287.x

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER IRELAND LTD  

Prosaposin acts as a neurotrophic factor, in addition to its role as the precursor protein for saposins A, B, C, and D, which are activators for specific sphingolipid hydrolases in lysosomes. In rats, the prosaposin gene generates two alternative splicing forms of mRNA: Pro + 9 containing a 9-base insertion and Pro + 0 without. The expression of these mRNAs changes after brain injury. We examined the expression patterns of the alternative splicing forms of prosaposin mRNA in the rat facial nerve nucleus for 52 days following facial nerve transection. Pro + 0 mRNA increased within 3 days of transection, peaked after 5-10 days, and remained significantly elevated for 21 days. In contrast, the expression of Pro + 9 mRNA was constant throughout the regenerative period. Prosaposin mRNA expression increased not only in facial motoneurons, but also in microglia during facial nerve regeneration. Our findings indicate that the saposin B domain of prosaposin, which is the domain affected by alternative splicing, plays an important role in both neurons and microglia during neuroregeneration. (c) 2007 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

DOI： 10.1016/j.neures.2007.09.010

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Prosaposin acts as a neurotrophic factor, in addition to its role as the precursor protein for saposins A, B, C, and D, which are activators for specific sphingolipid hydrolases in lysosomes. In rats, the prosaposin gene generates two alternative splicing forms of mRNA: Pro + 9 containing a 9-base insertion and Pro + 0 without. The expression of these mRNAs changes after brain injury. We examined the expression patterns of the alternative splicing forms of prosaposin mRNA in the rat facial nerve nucleus for 52 days following facial nerve transection. Pro + 0 mRNA increased within 3 days of transection, peaked after 5-10 days, and remained significantly elevated for 21 days. In contrast, the expression of Pro + 9 mRNA was constant throughout the regenerative period. Prosaposin mRNA expression increased not only in facial motoneurons, but also in microglia during facial nerve regeneration. Our findings indicate that the saposin B domain of prosaposin, which is the domain affected by alternative splicing, plays an important role in both neurons and microglia during neuroregeneration. © 2007 Elsevier Ireland Ltd and the Japan Neuroscience Society.

DOI： 10.1016/j.neures.2007.09.010

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DOI： 10.11172/keitaikinou2002.6.99

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Language：Japanese   Publisher：Japanese Physical Therapy Association (JPTA)  

We have introduced a human dissection course for the first year PT-OT students, where the students perform the dissection themselves, in other words "with their own scalpels", in 2004 supported by Ehime University. Prior to that, in 2003 we started human dissection training for faculty staffs at Ehime Juzen School of Allied Medical Professions. Based on questionnaires given to the students who participated in these courses, this study examined how students thought about and understand medical ethics related to the humanity, death and lives of the patients as well as of the students themselves. Analysis of the questionnaires lead to the conclusion that participation in human dissection gave students a considerable opportunity to think about medical ethics, which is necessary for those who work in hospitals and clinics. Further examination of the responses to the questionnaire suggested that the class work should utilize supplementary materials on human anatomy. In addition, many resources such as TV news, newspapers and recent movies, should be introduced into the small group- learning classes in order to help the students feel the realities related to ethical issues.

CiNii Books

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Publishing type：Research paper (scientific journal)  

Prosaposin is the precursor of four sphingolipid activator proteins (saposins A, B, C, and D) for lysosomal hydrolases and is abundant in the nervous system and muscle. In addition to its role as a precursor of saposins in lysosomes, intact prosaposin has neurotrophic effects in vivo or in vitro when supplied exogenously. We examined the distribution of prosaposin in the central and peripheral nervous systems and its intracellular distribution. Using a monospecific antisaposin D antibody that crossreacts with prosaposin but not with saposins A, B, or C, immunoblot experiments showed that both the central and peripheral nervous systems express unprocessed prosaposin and little saposin D. Using the antisaposin D antibodies, we demonstrated that prosaposin is abundant in almost all neurons of both the central and peripheral nervous systems, including autonomic nerves, as well as motor and sensory nerves. Immunoelectron microscopy using double staining with antisaposin D and anticathepsin D antibodies showed strong prosaposin immunoreactivity mainly in the lysosomal granules in the neurons in both the central and peripheral nervous systems. The expression of prosaposin mRNA, examined using in situ hybridization, was observed in these same neurons. Our results suggest that prosaposin is synthesized ubiquitously in neurons of both the central and peripheral nervous systems. © 2007 Springer-Verlag.

DOI： 10.1007/s00441-007-0464-9

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Language：English   Publishing type：Research paper (scientific journal)  

Prosaposin is the precursor of four sphingolipid activator proteins (saposins A, B, C, and D) for lysosomal hydrolases and is abundant in the nervous system and muscle. In addition to its role as a precursor of saposins in lysosomes, intact prosaposin has neurotrophic effects in vivo or in vitro when supplied exogenously. We examined the distribution ofprosaposin in the central and peripheral nervous systems and its intracellular distribution. Using a monospecific antisaposin D antibody that crossreacts with prosaposin but not with saposins A, B, or C, immunoblot experiments showed that both the central and peripheral nervous systems express unprocessed prosaposin and little saposin D. Using the antisaposin D antibodies, we demonstrated that prosaposin is abundant in almost all neurons of both the central and peripheral nervous systems, including autonomic nerves, as well as motor and sensory nerves. Immunoelectron microscopy using double staining with antisaposin D and anticathepsin D antibodies showed strong prosaposin immunoreactivity mainly in the lysosomal granules in the neurons in both the central and peripheral nervous systems. The expression of prosaposin mRNA, examined us

DOI： 10.1007/s00441-007-0464-9

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER IRELAND LTD  

Prosaposin, the precursor of the sphingolipid hydrolase activator proteins called saposins A, B, C, and D, is abundant in the nervous system and muscles. Besides its role as the precursor of saposins, prosaposin is reported to function as a neurotrophic factor, initiating neural differentiation and preventing neuronal cell death in vivo and in vitro. In this study, we examined the localization and synthesis of prosaposin in the rat cochlea. Intense prosaposin immunoreactivity was observed in the organ of Corti, stria vascularis, and spiral ganglion. In an immuno-electron microscopic study, prosaposin immunoreactivity was found mainly in lysosomal granules of the cells in these regions. In the lysosome, prosaposin does not always colocalize with cathepsin D, but was localized mainly in the dark area of the lysosome. Prosaposin mRNA was observed in these same regions. Our results suggest that prosaposin plays a role in homeostasis in the peripheral auditory system. (c) 2006 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

DOI： 10.1016/j.neures.2006.11.006

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER GMBH, URBAN & FISCHER VERLAG  

During our dissection of a Japanese elderly female cadaver, a double aortic arch with a deformed trachea was found in the cadaver. The ascending aorta was bifurcated to form the left (anterior) and right (posterior) aortic arches. Encircling and compressing the trachea and esophagus, they confluenced into the descending aorta. We concluded that it was a case of the double aortic arch forming a vascular ring. In the vascular ring the trachea was deformed. In addition, the left innominate vein coursed under the aortic arches (subaortic Left innominate vein, SLIV) and crossed the mediastinum posterior to the ascending aorta and anterior to the trachea. (c) 2006 Elsevier GmbH. All rights reserved.

DOI： 10.1016/j.aanat.2006.07.010

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Lectin binding patterns in the olfactory bulb of the Japanese common toad, Bufo japonicus, were examined using 21 types of lectin. Ten out of 21 lectins, WGA, s-WGA, LEL, STL, DBA, VVA, SJA, RCA-I, PNA, and PHA-L, stained the olfactory nerve, the glomeruli in the main olfactory bulb (MOB), the vomeronasal nerve, and the glomeruli in the accessory olfactory bulb (AOB). The binding patterns of LEL, STL, DBA, and PHA-L subdivided AOB glomeruli into rostral and caudal regions, where LEL, STL, and DBA stained the rostral region more intensely than the caudal region, and PHA-L had the opposite effect. Another lectin, BSL-I, stained both AOB glomeruli and the vomeronasal nerve, but not MOB glomeruli or the olfactory nerve. This is the first report of histological subdivision in the AOB of an amphibian, which suggests that the AOB development in Bufo may be unique.

DOI： 10.1007/s00429-006-0088-y

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-LISS  

An anomalous branch of the right coronary artery was found in a 71-year-old male cadaver with a right-sided aortic arch. The anomalous artery arose from the proximal portion of the right coronary artery and ran in a retroaortic course, before reaching the posterior wall of the heart. It was recognized as the right-sided variation of the circumflex coronary artery. The aortic arch had as branches the left common carotid, right common carotid, right subclavian, and left subclavian arteries, in that order, and the descending aorta was located in the right thorax. The left subclavian artery arose from a Kommerell's diverticulum and ran behind the esophagus, and the left-sided ligamentum arteriosum was also connected at the diverticulum. Therefore, the right aortic arch was classified as type N according to Adachi-Willimis-Nakagawa and type III-B1 in accordance with Stewart-Edwards. The Kornmerell's diverticulum in this case seemed to press on the posterior wall of the esophagus.

DOI： 10.1002/ca.20193

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：AMER SOC CLINICAL INVESTIGATION INC  

Muscle-specific kinase (MuSK) is critical for the synaptic clustering of nicotinic acetylcholine receptors (AChRs) and plays multiple roles in the organization and maintenance of neuromuscular junctions (NMJs). MuSK is activated by agrin, which is released from motoneurons, and induces AChR clustering at the postsynaptic membrane. Although autoantibodies against the ectodomain of MuSK have been found in a proportion of patients with generalized myasthenia gravis (MG), it is unclear whether MuSK autoantibodies are the causative agent of generalized MG. In the present study, rabbits immunized with MuSK ectodomain protein manifested MG-like muscle weakness with a reduction of AChR clustering at the NMJs. The autoantibodies activated MuSK and blocked AChR clustering induced by agrin or by mediators that do not activate MuSK. Thus MuSK autoantibodies rigorously inhibit AChR clustering mediated by multiple pathways, an outcome that broadens our general comprehension of the pathogenesis of MG.

DOI： 10.1172/JCI21545

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Language：English   Publisher：(一社)日本解剖学会  

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Language：Japanese   Publisher：(一社)日本解剖学会  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE  

We created chicks with spina bifida aperta (SBA) by incising the roof plate of the neural tube of embryos at Hamburger Hamilton stage 18 or 19. Incision over the length of three somites caused spina bifida occulta (SBO)-like malformation in 47% of the hatchlings. Incision over the length of five and seven somites caused SBA-like malformation in 100% of the hatchlings. The SBO chicks exhibited no symptoms, whereas the SBA chicks exhibited paralysis of a leg muscle and imbalance between all agonist and all antagonist leg muscles. Lesions in these SBA chicks were located in the spinal segments that give rise to motor neurons that innervated the dysfunctional muscles. Histological analysis revealed that there were fewer small spinal neurons (interneurons) at the site of the lesion in SBA chicks than in the normal chicks and that there was no such difference in the number of the large spinal neurons (motor neurons). Leg dysfunctions in this model of SBA may be attributable to the smaller number of interneurons in the spinal segments that contain motor neurons that innervate the dysfunctional muscle. This model may facilitate studies of the pathological mechanisms that lead to leg dysfunctions in SBA chicks. (c) 2005 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.expneurol.2005.09.00l

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An anomalous branch of the right coronary artery was found in a 71-year-old male cadaver with a right-sided aortic arch. The anomalous artery arose from the proximal portion of the right coronary artery and ran in a retroaortic course, before reaching the posterior wall of the heart. It was recognized as the right-sided variation of the circumflex coronary artery. The aortic arch had as branches the left common carotid, right common carotid, right subclavian, and left subclavian arteries, in that order, and the descending aorta was located in the right thorax. The left subclavian artery arose from a Kommerell's diverticulum and ran behind the esophagus, and the left-sided ligamentum arteriosum was also connected at the diverticulum. Therefore, the right aortic arch was classified as type N according to Adachi-Williams-Nakagawa and type III-B1 in accordance with Stewart-Edwards. The Kommerell's diverticulum in this case seemed to press on the posterior wall of the esophagus. © 2005 Wiley-Liss, Inc.

DOI： 10.1002/ca.20193

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-LISS  

Cajal's initial glomeruli (IG) and Dogiel's pericellular nests (PCNs) were first described from methylene blue preparations of healthy animal tissues around the beginning of the last century. Since that time, although many reports have been published concerning these structures, few have focused on their development and phylogeny in healthy animals. The aim of this study was to examine the phylogenetic development of the sensory neurons in Cajal's IG (also called axonal glomeruli) and Dogiel's PCNs in the dorsal root ganglion (DRG) of the healthy adult frog, chick, rat, and rabbit. The three-dimensional architecture of the neurons was observed in ganglia by scanning electron microscopy after removal of the connective tissue. The neurons in the DRG of fish are known to be bipolar, but DRG neurons in the species examined here were found to be pseudounipolar, with single stem processes. The proportion of neurons having IG or PCNs increased with increasing phylogenetic complexity in the species examined here. Cajal's initial glomeruli, the convolution of the stem process near the parent cell body: In frogs, the ganglia were small and the neuronal stem processes were very short and straight. In chicks, the stem processes were longer; sometimes very long, tortuous processes were observed. However, no neurons with typical IG were observed in either species. Typical IG were observed in rats and rabbits; their occurrence was much more frequent in rabbits. Pseudounipolarization, i.e., the transition from bipolar to pseudounipolar neurons, is thought to save space, limit the length of neuronal processes, and reduce conduction time. However, an explanation of the evolutionary advantage of the IG, which is formed by the excessive prolongation of the stem process, remains elusive. The cytological and electrophysiological importance of IG has been discussed. Dogiel's pericellular nests (PCNs), which resemble balls of yarn made of thin unmyelinated nerve fibers around DRG neurons, have been observed in the DRG of rats and rabbits, but not in frogs or chicks. This interesting structure shows not only ontogenetic development in healthy animals but also phylogenetic development among species. The nerve fibers in the PCNs were less than 1.2 mu m in diameter and had some varicosities. An immunohistochemical study using anti-tyrosine hydroxylase (TH) antibody revealed that some PCNs contain TH-positive nerve fibers and varicosities. Such TH-positive PCNs disappear after sympathectomy. These results suggest that the PCNs are made up of autonomic nerve fibers.

DOI： 10.1002/cne.20713

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Prosaposin is the precursor of saposins A, B, C and D, which are activators of sphingolipid hydrolases. In addition, unprocessed prosaposin functions as a neurotrophic factor in the central and peripheral nervous systems by acting to prevent neuronal apoptosis, to elongate neurites and to facilitate myelination. In this study, the expression pattern of prosaposin in the facial nerve nucleus after facial nerve transection was examined by immunohistochemistry and in situ hybridization. Prosaposin immunoreactivity in the neurons on the operated side facial nerve nucleus showed a biphasic pattern: it was significantly increased on day 3 after transection, decreased dramatically on day 7, started to increase gradually on day 14 and reached another peak on day 21 after transection. Significant increases in the levels of prosaposin mRNA were identified in the neurons on the operated side, suggesting that prosaposin was synthesized vigorously by the neurons themselves in the case of facial nerve transection. The diverse changes in prosaposin immunoreactivity during the process of facial nerve regeneration may reflect the diverse neurotrophic activities of prosaposin in facial motoneurons. © 2005 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

DOI： 10.1016/j.neures.2005.03.009

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE  

Adrenomedullin (AM) is postulated to exert organ-protective effects. It is expressed in the renal glomeruli, but its roles in the glomerular podocytes have been poorly elucidated. In the present study, we investigated the expression and regulation of AM in recently established conditionally immortalized mouse podocyte cell line in vitro and podocyte injury model in vivo. The cultured differentiated podocytes expressed AM mRNA and secreted measurable amount of AM. AM secretion from the podocytes was increased by H2O2, hypoxia, puromycin aminonucleoside (PAN), albumin overload, and TNF-alpha. Real-time RT-PCR analysis revealed that AM mRNA expression in the podocytes was enhanced by PAN and TNF-alpha, both of which were suppressed by mitochondrial antioxidants. Furthermore, AM expression was upregulated in the glomerular podocytes of PAN nephrosis rats. These results indicated that AM expression in the podocytes was upregulated by stimuli or condition relevant to podocyte injury, suggesting its potential role in podocyte pathophysiology. (c) 2005 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2005.02.142

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Language：Japanese   Publisher：愛媛十全医療学院  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ELSEVIER SCI IRELAND LTD  

In the developing central nervous system, apoptosis plays an important role in the normal organization of the neuronal circuit. The timing of neurogenesis, proliferation, and migration of the neurons in the developing olfactory bulb (OB) is well studied; however, the involvement of apoptosis in this process is not fully understood. In this study, we examined the changes in the distribution and the number of apoptotic cells in the rat OB during embryonic and postnatal periods, by using terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling (TUNEL) staining. Although the number of TUNEL-positive cells was relatively small during the embryonic period, it gradually increased after birth, and peaked on postnatal day 20 with statistical significance, especially in the granule cell layer of the main OB. This transient increase of TUNEL-positive cells on postnatal day 20 may be involved in a critical event during maturation of the OB. (C) 2004 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

DOI： 10.1016/j.neures.2004.07.001

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MOSBY, INC  

Background. Proliferation and apoptosis of liver cancer cells are closely related phenomena. We investigated the correlation between overexpression of Bcl-xL, an anti-apoptosis-related protein of the Bcl-2 family, and the clinical course of hepatocellular carcinoma (HCC).
Methods. Specimens from 7 HCC patients were used for Western blotting and immunoelectron microscopy tests. Samples from 33 HCC patients who had undergone hepatectomies were used for immunohistochemical staining. The degrees of expression of Bcl-xL and Ki-67, as an index of HCC mitosis severity, were each classified into 2 groups.
Results. With the use of Western blot analysis, enhanced immunoreactivity of Bcl-xL was found in cancerous specimens. Bcl-xL overexpression was found in cancer specimens in 21 of 33 patients (63.6%). The overall survival (P = .019) and disease-free survival (P = .030) rates of the group overexpressing Bcl-xL were definitely poorer. The Ki-67 higher labeling index LI &gt; 10) group had a poorer survival rate (P = .016). There were significant correlations between Bcl-xL and overall survival and disease-free survival. Multivariate analyses revealed that. Bcl-xL, tumor size, histologic portal invasion, and histologic metastatic foci were independent prognostic factors for overall survival and disease-free survival.
Conclusions. These results showed Bcl-xL in HCC specimens, suggesting that Bcl-xL was a significant Prognostic factor for disease progression in human HCC.

DOI： 10.1016/j.surg.2003.11.015

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Language：Japanese   Publishing type：Research paper (bulletin of university, research institution)  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KARGER  

Background/Aims: Megalin is a multiligand endocytic receptor expressed in a number of epithelia. In the Lewis rat kidney, podocytes, as well as proximal tubule cells, express megalin that acts as a pathogenic antigen for Heymann nephritis (HN), an experimental model of membranous nephropathy. To obtain a tool to investigate the molecular mechanisms of megalin-mediated endocytosis and the pathogenesis of HN, we examined whether a differentiation-inducible mouse podocyte cell line expressed endocytically active megalin. Methods: Immunofluorescence and immunoprecipitation analyses with an anti-rat megalin antibody were carried out to investigate whether megalin was expressed in the differentiated and undifferentiated podocytes. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis was performed to elucidate whether the cells synthesize megalin mRNA. I-125-labeled receptor-associated protein (RAP), an endocytic ligand for megalin, was used for cellular internalization and degradation assays. Results: Immunofluorescence, immunoprecipitation and RT-PCR analyses revealed that megalin was synthesized in both differentiated and undifferentiated cells and localized to the cell surfaces. Effective endocytosis of RAP via megalin was shown under the differentiated condition. Conclusion: Endocytically active megalin is expressed in differentiation-induced cultured podocytes. This cell line could be a useful tool for studies on megalin-mediated endocytosis and the pathogenesis of HN. Copyright (C) 2004 S. Karger AG, Basel.

DOI： 10.1159/000076404

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DOI： 10.1159/000078406

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KARGER  

Background: Podocytes, renal glomerular visceral epithelial cells, have two kinds of processes, namely major processes containing microtubules (MTs) and foot processes with actin filaments (AFs). The present study investigated how MTs are organized by the Rho-ROCK signal transduction pathway during process formation of podocytes. Method: After induction of differentiation, podocytes of the conditionally immortalized mouse cell line were treated with Y-27632, a specific inhibitor of ROCK, and exoenzyme C3, an inhibitor of RhoA, as well as with forskolin whose effects include inhibition of RhoA, in order to inhibit the Rho-ROCK pathway. Results: Inhibition of ROCK significantly enhanced the formation of thick processes containing MT bundles. Y27632 promoted process formation even in the presence of latrunculin A which disrupts AFs, strongly suggesting that ROCK directly regulates MT assembly. Treatment with Y-27632 increased MT stability, and stabilized MTs preferentially localized in podocyte processes. Moreover, when treated with a combination of Y-27632 and forskolin, and with Y-27632 and C3 as well, podocytes developed not only MT-based thick processes but also AF-based thin projections. Conclusions: These data indicate a contribution of ROCK in MT organization to promote podocyte process formation, although it was originally thought to regulate AF assembly. AF-based thin projections seem to be induced mainly by inhibition of RhoA and ROCK. The present study reveals a significant role of the Rho-ROCK signal pathway in the reorganization of both MTs and AFs during process formation of podocytes. Copyright (C) 2004 S. Karger AG, Basel.

DOI： 10.1159/000078406

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：LIPPINCOTT WILLIAMS & WILKINS  

IFN-inducible protein-10 (IP-10/CXCL10) is a potent chemoattractant for activated T lymphocytes and was recently reported to have several additional biologic activities. In this study, the expression and the function in normal glomeruli and in Thy1.1 glomerulonephritis (GN) were investigated. The expression of IP-10 was detected in normal rat glomeruli mainly in the podocyte. The expression of IP-10 was also detected on the cultured podocyte. The IP-10 expression was elevated at the early phase of Thyl.1 GN. The double staining immunofluorescence study clearly demonstrated that the elevated expression of IP-10 was mostly detected in the podocyte and very partly in mesangial area. A receptor for IP-10, CXCR3, showed similar expression patterns to that of IP-10. Expressions of neither of IP-10 nor of CXCR3 were detected on the inflammatory cells. For elucidating the role of IP-10, the blocking study was carried out with monoclonal anti-IP-10 antibody. The monoclonal anti-IP-10 antibody treatment decreased the expression of IP-10 and podocyte associated proteins such as nephrin and podocin that are reported to be essential for maintaining the podocyte function (IP-10, 53.0% to control; nephrin, 43.5%; podocin, 60.4%). The findings indicated that the anti-IP-10 treatment disturbed the podocyte function. The anti-IP-10 treatment given to the rats with Thy1.1 nephritis exacerbated proteinuria, mesangiolysis, and matrix expansion. Collectively, the findings indicated that IP-10 plays a role in maintaining the podocyte function. Also, the findings suggested that anti-IP-10 treatment exacerbated the glomerular alterations in Thyl.1 GN by disturbing the podocyte function.

DOI： 10.1097/01.ASN.0000097371.64671.65

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：TAYLOR & FRANCIS LTD  

We previously identified a 65-kDa protein (p65) that was phosphorylated in activated macrophages. It has turned out to be a murine homologue of human L-plastin, which was identified as a novel protein in human cancer cells. p65/L-plastin is characterized by a series of Ca2+-, calmodulin-, and actin-binding domains, and is thought to play a crucial role in leukocytes and cancer cells. We have expressed a recombinant (r) p65/L-plastin in Escherichia coli that binds to beta-actin and prepared high-titer antibodies using large amounts of the protein as immunogen. Anti-rp65/L-plastin antibodies recognize native p65/L-plastin as well as rp65/L-plastin and have enabled us to detect the fine structures of intracellular p65/L-plastin, and it was found that its localization was extensively changed by stimulation with bacterial components. We further developed an enzyme-linked immunosorbent assay system and a flow cytometry method using these reagents, which made it possible to measure antibodies, including autoantibodies, against p65/L-plastin and to evaluate the maturation-dependent expression of the protein in leukocytes.

DOI： 10.1271/bbb.67.1368

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

Recently, beta-catenin has been reported to control the expression of morphogenetic genes through the Wnt signaling pathway in invertebrate embryogenesis. In this study, the distribution pattern of beta-catenin during starfish embryogenesis was investigated using immunohistochemistry. In 16-cell stage embryos, beta-catenin began to accumulate in some nuclei at the vegetal pole. During the early cleavage stage, the cells expressing nuclear beta-catenin increased in number in the vegetal pole region of the embryos, and the beta-catenin signal increased in intensity in each nucleus. At the blastula stage, signal for beta-catenin was also found in the cytoplasm of the cells with nuclear beta-catenin. At the vegetal plate stage, almost all vegetal plate cells expressed beta-catenin in both the nucleus and cytoplasm. When the embryos developed to early gastrulae, cells with nuclear beta-catenin were restricted to the archenteron tip, and the signal gradually faded in later stages. The localization and temporal change of beta-catenin expression suggests that beta-catenin has a pivotal role in archenteron formation in starfish embryos.

DOI： 10.1034/j.1600-0854.2004.00681.x

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER-VERLAG  

Expression patterns of glycoconjugates were examined by lectin histochemistry in the nasal cavity of the Japanese red-bellied newt, Cynops pyrrhogaster. Its nasal cavity consisted of two components, a flattened chamber, which was the main nasal chamber (MNC), and a lateral diverticulum called the lateral nasal sinus (LNS), which communicated medially with the MNC. The MNC was lined with the olfactory epithelium (OE), while the diverticulum constituting the LNS was lined with the vomeronasal epithelium (VNE). Nasal glands were observed beneath the OE but not beneath the VNE. In addition, a secretory epithelium was revealed on the dorsal boundary between the MNC and the LNS, which we refer to as the boundary secretory epithelium (BSE) in this study. The BSE seemed to play an important role in the construction of the mucous composition of the VNE. Among 21 lectins used in this study, DBA, SBA and Jacalin showed different staining patterns between the OE and the VNE. DBA staining showed remarkable differences between the OE and the VNE; there was intense staining, in the free border and the supporting cells of the VNE, whereas there was no staining or weak staining in the cells of the OE. SBA and Jacalin showed different stainings in the receptor neurons for the OE and the VNE. Furthermore, UEA-I and Con A showed different stainings for the nasal glands. UEA-I showed intense staining in the BSE and in the nasal glands located in the ventral wall of the MNC (VNG), whereas Con A showed intense staining in the BSE and in the nasal glands located in the dorsal and medial wall of the MNC (DMNG). The DMNG were observed to send their excretory ducts into the OE, whereas no excretory ducts were observed from the VNG to the OE or the VNE. These results suggested that the secretion by the supporting cells as well as the BSE and the DMNG establishes that there are heterogeneous mucous environments in the OE and the VNE, although both epithelia are situated in the same nasal cavity.

DOI： 10.1007/s00429-002-0306-1

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DOI： 10.1002/ca.10057

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Language：Japanese   Publishing type：Research paper (bulletin of university, research institution)  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE  

In 1995, we developed an animal model of transient homolateral facial nerve paralysis by inoculating Herpes simplex virus type 1 (HSV-1) into the auricle of mice. This study examined the mechanism of facial nerve paralysis in this model histopathologically. Using the immunofluorescence technique with anti-HSV-1 antibody, the time course of viral spread and the site of viral replication were investigated over the entire course of the facial nerve. Furthermore, viral replication and nerve degeneration at the site of viral replication were observed by electron microscopy. On the 7th day after inoculation, facial paralysis was observed in more than 60% of mice. Immunofluorescence study revealed HSV-1 in the geniculate ganglion, the descending root, and the facial nucleus at this stage. On the 9th day, the descending root in the sections stained with osmium. looked pale, because prominent demyelination had occurred in this region; electron micrographs showed many degenerated oligodendrocytes and large naked axons. In contrast, the facial nucleus neurons showed no remarkable degeneration, despite HSV-1 particles in their cytoplasm. From these findings, we concluded that facial nerve paralysis in this model is caused mainly by facial nerve demyelination in the descending root. (C) 2002 Elsevier Science (USA).

DOI： 10.1006/exnr.2002.8035

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Language：Japanese   Publisher：愛媛医学会  

ヒト顔面神経麻痺の再生過程を知ることを目的に,ラットのモデルを用い,顔面神経切断後の顔面神経細胞内のプロサポシンの増減を免疫組織化学的手法により検討した.ラットの右側顔面神経の切断を行い,正常側の免疫反応と比較した.その結果,顔面神経核内のプロサポシン免疫反応は術後3日に一過性に増加し,7日後に著しく低下し,14日後に回復傾向を示し,21日後に再増加のピークを示した.術後3日の免疫反応は,障害された神経細胞がプロサポシンを産出し,自らを保護する働きによると判断した.術後21日は神経軸索の再形成期と一致しており,増加したプロサポシンが髄鞘再形成を刺激すると推測した.7日後,14日後は,軸索膜損傷後の神経細胞内タンパク合成能の低下によりプロサポシン産生が減少したため,反応が低下したと判断した

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：PROFESSOR D A SPANDIDOS  

It is known that proliferation and apoptosis are closely related phenomena in liver cancer cells. In this study, using surgical specimens from 42 patients with hepatocellular carcinoma (HCC), we investigated the expression and localization of Bcl-xL, an antiapoptosis-related protein of the Bcl-2 family. Using Western blotting, Bcl-xL expression was detected in both cancerous and non-cancerous specimens from all of the HCC patients, and elevated Bcl-xL levels were found in cancerous specimens from two thirds of the patients. In normal human liver specimens, Bcl-xL was found mainly in the cytoplasm of hepatocytes, although it was also found in the cytoplasm of bile duct cells in Glisson's capsule by immunohistochemical staining. To the best of our knowledge, this is the first report of Bcl-xL overexpression and localization in HCC specimens. Bcl-xL was found not only in the cytoplasm of HCC cells, but also in the nuclei of some HCC cells, suggesting that Bcl-xL is involved in the progression of HCC cells in vivo.

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：フレグランスジャーナル社  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KLUWER ACADEMIC PUBL  

This study presents the first direct evidence for herpes simplex virus type 1 (HSV-1) infection in the neurons of the vestibular ganglion. Although many investigators have reported electron microscopic evidence of HSV-1 infection in sensory ganglia, HSV-1 infection in the vestibular ganglion has not been described. Vestibular ganglion neurons have a unique structure, with a loose myelin sheath instead of the satellite cell sheath that is seen in other ganglia. This loose myelin is slightly different from compact myelin which is known as too tight for HSV-1 to penetrate. The role of loose myelin in terms of HSV-1 infection is completely unknown. Therefore, in an attempt to evaluate the role of loose myelin in HSV-1 infection, we looked for HSV-1 particles, or any effects mediated by HSV-1, in the vestibular ganglion as compared with the geniculate ganglion. At the light microscopic level, some neurons with vacuolar changes were observed, mainly in the distal portion of the vestibular ganglion where the communicating branch from the geniculate ganglion enters. At the electron microscopic level, vacuoles, dilated rough endoplasmic reticulum and Golgi vesicles occupied by virus were observed in both ganglia neurons. In contrast, viral infections in Schwann and satellite cells were observed only in the geniculate ganglion, but not in the vestibular ganglion. These results suggest that loose myelin is an important barrier to HSV-1 infection, and it must play an important role in the prevention of viral spread from infected neurons to other cells.

DOI： 10.1023/A:1016577700280

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER-VERLAG  

Podocytes possess major processes containing microtubules (MTs) and intermediate filaments and foot processes containing actin filaments (AFs) as core cytoskeletal elements. Although the importance of these cytoskeletal elements for maintaining podocyte processes was previously shown, so far no data are available concerning the developmental regulation of podocyte process formation. A conditionally immortalized mouse podocyte cell line, which can be induced to develop processes similar to those found in vivo, was treated with various reagents to disrupt cytoskeletal elements or to inhibit protein phosphatases, MTs colocalized with vimentin intermediate filaments but not with AFs. After AF disassembly, major processes were maintained, whereas after depolymerization of MTs, podocytes lost their processes, rounded up, and maintained only actin-based pe ripheral projections. Suppression of MT elongation by nanomolar vinblastine or inhibition of serine/threonine phosphatase PP2A with okadaic acid abolished process formation. PP2A was expressed in undifferentiated but not in differentiated podocytes. One- and two-dimensional western blot analyses revealed a dose-dependent increase in serine/threonine phosphorylation after okadaic acid treatment. Hence, morphogenetic activity of MTs induces podocyte process formation via serine/threonine protein dephosphorylation by PP2A. These results may open new avenues for understanding the signaling mechanism underlying podocyte cytoskeleton alterations during development and in glomerular diseases.

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：EDITRICE IL SEDICESIMO  

Morphogenetic effects of various extracellular matrix proteins on the renal podocyte were investigated using the conditionally immortalized podocyte cell line. Podocytes were plated on glass coverslips and coated with the following matrix proteins: laminin-10/11, laminin-1, fibronectin, collagen type IV, collagen type L Three hours after plating, podocytes on laminins developed prominent processes, while those on other matrix proteins started to elongate processes after two days. Vinculin-immunolabeling showed that podocytes plated on laminins possessed thin rod-shaped focal contacts, whereas those on fibronectin showed large dot-shaped focal contacts. Inhibition of serine/threonine protein kinases induced podocyte process formation in an extracellular matrix-independent manner. The present study reveals the significance of laminin on podocyte morphogenesis in vitro, and shows that different extracellular matrix proteins trigger different intracellular signals governing podocyte morphogenesis. Taken together with our previous studies, podocyte process formation is thought to be regulated by protein Ser/Thr phosphorylation.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ACADEMIC PRESS INC  

The identity of two microtubule-associated proteins, MAP3 and MAP4, was verified both immunologically and biochemically. MAP3 was enriched from the heat-stable fraction of rat brain extracts by reverse-phase HPLC and preparative two-dimensional gel electrophoresis, Both MAP3 and MAP4 antibodies reacted with the corresponding spots on two-dimensional Western blots. Amino acid sequences of internal peptides derived from rat MAP3 matched with corresponding sequence stretches of mouse MAP4. In the kidney cortex, the MAP3 antibody stained not only glomerular podocytes but also interstitial cells. This distribution pattern of MAP3 is identical to that of MAP4 reported previously. These results indicate that MAP3 and MAP4 are identical. (C) 2000 Academic Press.

DOI： 10.1006/bbrc.2000.2126

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Language：Japanese   Publisher：(一社)日本解剖学会  

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Language：English   Publishing type：Research paper (scientific journal)  

The development of sensory ganglion neurons and satellite cells examined by scanning electron microscopy after removal of the connective tissue is reviewed. Sensory neurons are bipolar at early stages of development and later became pseudounipolar. This maturation event starts earlier but proceeds more slowly in chick than in rat embryos. These may due to the difference in the extent and intimacy of satellite cell investments between these two animal species. The neuronal perikaryal projections are observed by scanning electron microscopy after removal of the connective tissue and satellite cells. The morphometric analysis reveals that perikaryal projections are mom numerous on the surface of mature pseudounipolar neurons than on that of premature bipolar neurons; they increase in number as the neuronal cell bodies grow larger. This may support the hypothesis that perikaiγal projections are structural devices for increasing the neuron-satellite interface and for improving the efficiency of metabolic exchange between these two cell types. The important role of satellite cells in neuronal maturation is discussed. © The Bulgarian-American Center.

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Language：Japanese   Publishing type：Research paper (scientific journal)  

Microtubular cytoskeletons play a crucial role in the morphogenesis of process-bearing cells, such as the neuron and the renal glomerular podocyte. Microtubules are bundled and stabilized by various microtubule-associated proteins, providing a mechanical basis to maintain the deviated morphology of cell processes. To support the process morphology, microtubules are also associated with other cytoskeletal elements such as actin and intermediate filaments. The microtubular polarity is uniformly plus-end-distal in neuronal axons, whereas in dendrites as well as in podocytes, the polarity is revealed to be non-uniform (i.e., both plus-end-distal and minus-end-distal microtubules are present in cell processes). Recently, this non-uniformity is reported to be established by a microtubule-dependent motor protein. Motor proteins are capable to drive the intracellular transport of cytoskeletal elements in addition to that of membrane vesicles. It is still an open question whether cytoskeletal elements are transported along cell processes as subunits or as polymers.

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Podocytes are unique cells that are decisively involved in glomerular filtration. They are equipped with a complex process system consisting of major processes and foot processes whose function is insufficiently understood (Mundel, P., and W. Kriz. 1995. Anat. Embryol. 192:385-397). The major processes of podocytes contain a microtubular cytoskeleton. Taking advantage of a recently established cell culture system for podocytes with preserved ability to form processes (Mundel, P., J. Reiser, A. Zuniga Mejia Borja, H. Pavenstadt, G.R. Davidson, W. Kriz, and R. Zeller. 1997b. Exp. Cell Res. 36:248-258), we studied the functional significance of the microtubular system in major processes. The following data were obtained: (a) Microtubules (MTs) in podocytes show a nonuniform polarity as revealed by hook-decoration. (b) CHO1/ MKLP1, a kinesin-like motor protein, is associated with MTs in podocytes. (c) Treatment of differentiating podocytes with CHO1/MKLP1 antisense oligonucleotides abolished the formation of processes and the nonuniform polarity of MTs. (d) During the recovery from taxol treatment, taxol-stabilized (nocodazoleresistant) MT fragments were distributed in the cell periphery along newly assembled nocodazole-sensitive MTs. A similar distribution pattern of CHO1/MKLP1 was found under these circumstances, indicating its association with MTs. (e) In the recovery phase after complete depolymerization, MTs reassembled exclusively at centrosomes. Taken together, these findings lead to the conclusion that the nonuniform MT polarity in podocytes established by CHO1/MKLP1 is necessary for process formation.

DOI： 10.1083/jcb.143.7.1961

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Language：English   Publisher：SPRINGER  

This review discusses the role of microtubules in the formation of processes from neuronal and non-neuronal cells. In elongating axons of the neuron, tubulin molecules are transported toward the end of pre-existing microtubules, which may be nucleated at the centrosome, via a mechanism called slow axonal flow. Two different hypotheses are presented to explain this mechanism; the transport of soluble monomers and/or oligomers versus the transport of polymerized microtubules. The majority of tubulin seems to be transported as small oligomers as shown by the data presented so far. Alternatively, an active transport of polymerized microtubules driven by microtubule-based motor proteins is postulated as being responsible for the non-uniform polarity of microtubule bundles in dendrites of the neuron. Microtubule-associated proteins (MAPs) play a crucial role in stabilizing the microtubular arrays, whereas the non-uniform polarity of microtubules may be established with the aid of microtubule-based motor proteins. The signals activating centrosomal proteins and MAPs, resulting in process formation, include phosphorylation and dephosphorylation of these proteins. Not only neuronal cells, but also renal glomerular podocytes develop prominent cell processes equipped with well-organized microtubular cytoskeletons, and intermediate and actin filaments. A novel cell culture system for podocytes, in which process formation can be induced, should provide further evidence that microtubules play a pivotal role in process formation of non-neuronal cells.

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DOI： 10.1007/BF02479929

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：Japanese Association of Anatomists  

Sensory ganglion neurons in higher vertebrates are unique in that they are pseudounipolar with a single stem process that divides at some distance from the cell body into central and peripheral processes. In the early stages of development, these neurons are bipolar but later they became pseudounipolar. This developmental process of sensory ganglion neurons with satellite cells was examined by scanning electron microscopy following removal of connective tissue. This pseudo-unipolarization began earlier but proceeded at a slower rate in chick than in rat embryos. This difference may due to the difference found in the extent and intimacy of satellite cell investments in these two animals, which was due to the fact that sensory neurons undergo pseudo-unipolarization only in the presence of satellite cells in vitro. The neuronal perikaryal projections were observed by scanning electron microscopy after removal of connective tissue and satellite cells. Morphometric analysis revealed that perikaryal projections were more numerous on the surface of mature pseudounipolar neurons than on the surface of premature bipolar neurons, and that the number of projections increased as the neuronal cell bodies grew larger. This may support the hypothesis that perikaryal projections are structural devices for increasing the neuron-satellite interface and for improving the efficiency of metabolic exchange between these two cell types. These results suggest that satellite cells play an important role in neuronal maturation.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER VERLAG  

The distribution of aortic intimal smooth muscle cells in the normal rat during postnatal development was studied by electron microscopy and by staining with fluorescence-labeled phalloidin. The phenotypes of intimal and medial smooth muscle cells were almost identical at first; however, during development, the former remained synthetic, whereas the latter became contractile. Confocal laser scanning microscopy was utilized to observe intimal and medial cells separately. Intimal smooth muscle cells were rarely observed in neonatal rats, but appeared by 10 days of age and increased during postnatal development. A combination of confocal and conventional fluorescent microscopy clearly demonstrated that the intimal smooth muscle cells were preferentially distributed in: (1) the right-lateral and dorsal wall of the upper thoracic aorta, (2) the left-lateral and ventral wall of distal two-thirds of the descending aorta, and (3) the downstream side of branch orifices. Intimal smooth muscle cells in group (1) were oriented randomly, whereas most in group (2) ran longitudinally. Intimal smooth muscle cells at branches in group (3) ran obliquely from the edges at the downstream side in an upstream direction. They tended to accumulate in regions of the aortic wall considered to be under high tensile stress.

DOI： 10.1007/s004410050894

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：BLACKWELL SCIENCE INC  

The sequence of morphological changes during foot process effacement in acute puromycin aminonucleoside (PAN) nephrosis was examined by means of NaOH maceration and freeze cracking for scanning electron microscopy (SEM). The micrographs of SEM and those of transmission electron microscopy (TEM) were quantitatively analyzed by computerized morphometry, and were correlated with renal function. On day 2 after PAN injection, the slit length was moderately decreased by both shortening and degradation of the foot processes. On day 4, membrane-bounded vesicles were scattered in the lamina rara externa. During foot process effacement, the basal surface of podocytes developed palm-like domains that represented the cytoplasmic areas between interdigitation. The decrease in the length of podocyte cell borders paralleled the decrease of 2 C-hour creatinine clearance. The developement of the palm-like domains on the basal aspects of podocytes estimated by distance class analysis closely correlated with the sudden onset of proteinuria. We conclude that foot process effacement in PAN nephrosis caused by the retraction and degradation of foot processes leads to the development of palm-like domains, which is correlated with podocyte detachment as well as massive proteinuria.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER VERLAG  

Distribution of glomerular basement membrane (GEM) outpockets and dimensional growth of glomeruli were studied in the maturing stage of rat glomeruli after completion of nephrogenesis. We observed the postnatal rat glomeruli from 5 to 60 days of age by transmission electron microscopy and estimated the structural development of glomeruli by computerized morphometry. On day 5, the GEM was double in structure, possessing an epithelial and endothelial lamina densa. After day 10, the lamina densa of the GEM was single and sent branches toward the epithelial side making outpockets. There is no change in the distributional pattern of the outpockets, at least from day 10 to day 60, although they decreased considerably in number between days 20 and 40. They were found almost exclusively on the peripheral surface of the glomerulus. The rat glomeruli increased in volume constantly in this period, and the capillary volume increased more significantly than the mesangial volume. The GEM surface area increased in parallel with the glomerular tuft volume. The growing mode of capillaries was different before and after day 40; namely before day 40 elongation was predominant, whereas after day 40 widening was more pronounced. These results indicate that if the outpockets are the other site of GEM assembly after fusion of double basement membranes, the GEM must be redistributed from the peripheral to the paramesangial site to enable elongation and branch formation of capillaries during the growth of glomeruli.

DOI： 10.1007/s004670050140

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：BLACKWELL SCIENCE INC  

The three-dimensional structure of the mesangium was analyzed by means of reconstruction from serial semithin and ultrathin sections of the rat glomerulus. The mesangial domains traced on light micrographs of semithin sections were transferred to styrene models, which were stacked up to reconstruct the whole mesangium. The reconstructed mesangium was tree-like in shape nd was divided into three lobes that were connected to the vascular pole by a slender neck. The glomerulus contained no islets of mesangium which were not connected to the vascular pole. The mesangium contained mesangial loops that were penetrated by capillaries. Reliability of the findings on the mesangial loops was ascertained by various methods including reconstruction of part of the mesangium from ultrathin sections. Electron microscopic observations revealed that the arms of the mesangial loops were frequently very slender and consisted of mesangial cell processes containing prominent bundles of actin filaments. The mesangial loops were distributed evenly within the mesangium. Considering previous reports showing about 400 capillary the rat glomerulus as well as the present findings, we the mesangial loops may change the distribution of intraglomerular blood flow bq dynamic contraction of the mesangial cells, in or serve a's an additional safety device to prevent the expansion of glomerular capillaries.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：ROCKEFELLER UNIV PRESS  

We investigated immunohistochemical localization of V2 vasopressin receptor along the nephron using a specific polyclonal antibody. Staining was observed in some of thick ascending limbs and all of principal and inner medullary collecting duct (IMCD) cells. Not only basolateral but also luminal membrane was stained in collecting ducts, especially in terminal IMCD (tIMCD).
To learn the functional role of luminal V2 receptor in tIMCD, we studied the luminal effects of arginine vasopressin (AVP) on osmotic water permeability (Pf), urea permeability (Pu), and cAMP accumulation using isolated perfused rat tIMCD. In the absence of bath AVP, luminal AVP caused a small increase in cAMP accumulation, Pf and Pu, confirming the presence of V2 receptor in the lumen of tIMCD. In contrast, luminal AVP inhibited Pf and Pu by 30-65% in the presence of bath AVP by decreasing cAMP accumulation via V1a or oxytocin receptors and by an unknown mechanism via V2 receptors in the luminal membrane of tIMCD.
These data show that V2 receptors are localized not only in the basolateral membrane but also in the luminal membrane of the distal nephron, Luminal AVP acts as a negative feedback system upon the basolateral action of AVP in tIMCD.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER VERLAG  

The structure of the normal pulmonary arteries in the rat was studied with light and electron microscopy after use of a newly devised technique of perfusion fixation and tissue preparation. We distinguished two main types of artery in the rat lung on the basis of the structure of the media, an elastic artery and a muscular artery. The elastic artery was characterized by an abundance of extracellular matrix in the media and by an oblique arrangement of smooth muscle cells to connect neighboring elastic laminae. It was subdivided into two segments, a classical elastic and a transitional elastic segment. The muscular artery was distinguished by a paucity of extracellular matrix in the media and by a circumferential arrangement of smooth muscle cells (or pericytes) enclosing the lumina, and was subdivided into four segments, a thick muscular, an ordinary muscular, a partially muscular and a nonmuscular segment. The smooth muscle cells in the muscular artery contained well-developed microfilament bundles compared with those in the elastic artery. Structural differences in smooth muscle cells and in extracellular matrix in the media between the elastic and muscular arteries may re fleet the functional heterogeneity of pulmonary arteries in response to hypoxic pulmonary vasoconstriction and to vasoactive substances such as endothelium-derived relaxing and hyperpolarizing factors, and endothelin.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Nature Publishing Group  

The contractility and distensibility of renal arterioles are important in the regulation of glomerular filtration. However, little is known regarding the characteristics of contractile proteins in these arterioles. Recently it was demonstrated that vascular smooth muscles contain two types of myosin heavy chain (MHC) isoforms, SM1 and SM2, which are unique molecular markers of smooth muscle cell phenotypes. SM1 is constitutively expressed in all types of smooth muscles, whereas SM2 exists only in mature smooth muscles. We characterized the expression of MHC isoforms as well as the ultrastructural myofilament assembly of renal arteriolar smooth muscles in human, rat and rabbit by immunohistochemical techniques. SM1 and α-smooth muscle actin were localized in both the preglomerular vessels (including the afferent arterioles) and efferent arterioles, whereas SM2 was present only in the preglomerular vessels. Renin-producing cells in the afferent arterioles (juxtaglomerular granular cells, JG cells) were positive for α-smooth muscle actin but negative for SM2. When renin synthesis was stimulated, the more proximal afferent arteriolar smooth muscles turned renin-positive and SM2 disappeared. Glomerular mesangial cells did not show immunoreactivities for SM1, SM2 or α-smooth muscle actin. The difference in MHC isoform expression in these arterioles was also reflected by ultrastructures
the afferent arteriolar smooth muscles contained abundant myofilaments including thick filaments, whereas the efferent arteriolar smooth muscles had a few myofilaments composed only of thin microfilaments. The JG cells displayed a myofilament assembly similar to that in the efferent arteriolar smooth muscles. We conclude from these observations that smooth muscles in pre- and postglomerular arterioles, the glomerular mesangial cells and JG cells differ in phenotypes, suggesting that they may have different contractile properties which may be critically involved in the regulation of glomerular filtration.

DOI： 10.1038/ki.1995.305

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER VERLAG  

Postnatal change in the distribution of actin filaments in endothelial cells was studied in the rat aorta by use of rhodamine-phalloidin staining and confocal laser scanning microscopy. Endothelial cells of the rat aorta possessed two populations of actin filament bundles, namely, peripheral bands at the cell border and stress fibers running longitudinally in the cytoplasm. Aortic endothelial cells of the neonatal rat contained only stress fibers, whereas those of the 10-day-old rat developed both peripheral bands and stress fibers. After 20 days of age, aortic endothelial cells had predominantly peripheral bands with occasional stress fibers around the branch orifices. During postnatal development the length density of stress fibers in aortic endothelial cells decreased, whereas individual stress fibers in endothelial cells were shortened. Electron-microscopic observation revealed that the high intercellular boundaries of aortic endothelial cells at birth decreased in height and developed cytoplasmic interdigitations after 20 days of age. The occurrence of peripheral bands at the cell border is thought to be closely related to formation of cytoplasmic interdigitation which strengthens the mechanical connection between endothelial cells against increasing transmural pressure. Expression of stress fibers in aortic endothelial cells of the neonatal rat is supposed to be affected by longitudinal elongation of the developing aorta, whereas their postnatal decrease is thought to be correlated with the change of fluid shear stress loaded on the aortic endothelium.

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Language：English   Publisher：WILLIAMS & WILKINS  

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：KARGER  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WISSENSCHAFTLICHE VERLAG MBH  

The distribution of actin filaments (AFs) in arterial and arteriolar endothelial cells (ECs) was examined in situ by staining with rhodamine-phalloidin in the rat kidney after perfusion-fixation under physiological pressure. ECs possessed two populations of AF bundles, namely, peripheral bands (PBs) at the cell border and stress fibers (SFs) in the cytoplasm. The distribution of AFs was heterogeneous and exhibited three patterns in the arterial tree. In large arteries (&gt; 120 mum in luminal diameter) ECs contained predominantly PBs in addition to occasional SFs mainly in the upstream side of cells (PB-pattern). In arteries with intermediate diameters (30-120 mum), both PBs and SFs were conspicuous (PB-SF-pattern). In small arteries and arterioles (&lt;45 mum), SFs were prominent and PBs were rare (SF-pattern). In electron micrographs, we found that PBs were attached not to the lateral but to the basal plasma membrane via electron-dense materials and that SFs included two subpopulations, namely, thick basal and thin apical SFs. Morphometric analysis revealed that the length density of PBs was higher in segments with the PB-pattern than in those with the PB-SF-pattern. The length density of SFs was significantly higher in the SF-pattern than in the PB-SF-pattern, whereas there was little difference in this parameter among vessel segments with the SF-pattern. We suggest that PBs and SFs are fundamentally structures that anchor ECs on the substratum, and are differentiated according to their different mechanical burdens.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SPRINGER VERLAG  

The structure of the intracellular actin filaments and the extracellular matrices was studied in the distal interlobular arteries in the rat kidney, employing three different morphological techniques, including rhodamin-phalloidin staining of cryosections, resorcin-fuchsin staining of paraffin sections, and a cold dehydration procedure for electron microscopy. The endothelial cells possess longitudinally running stress fibers. The inner elastic layer is composed of meshworks of elastic fibers encompassing numerous pores. The smooth muscle cells containing abundant actin filaments are arranged circumferentially around the vascular axis. The endothelial stress fibers are found mainly in the basal half of the endothelial cells, and anchor onto the basal cell membranes. The elastic meshworks send off longitudinal branch fibers to contact the endothelial cell membranes at the anchoring sites of stress fibers. In addition circumferential branches run toward the smooth muscle cells. The functional significance of the intracellular contractile apparatus and the extracellular tensile component in small arteries was discussed.

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-LISS  

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)  

DOI： 10.24489/jjphe.2019-020

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Language：Japanese   Publisher：(一社)日本医学教育学会  

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Language：Japanese   Publisher：(一社)日本病理学会  

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Authorship：Lead author   Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (bulletin of university, research institution)  

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)  

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Language：Japanese   Publisher：(一社)日本医学教育学会  

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)   Publisher：(公社)愛媛県理学療法士会  

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)   Publisher：愛媛医学会  

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)   Publisher：愛媛医学会  

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Authorship：Lead author,　Corresponding author   Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)   Publisher：愛媛医学会  

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Authorship：Lead author   Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)   Publisher：(株)医学書院  

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Other Link： http://search.jamas.or.jp/link/ui/2016059209

Authorship：Lead author   Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)  

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DOI： 10.1016/j.neures.2006.11.006

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：WILEY-BLACKWELL  

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Language：Japanese   Publisher：(公社)日本薬理学会  

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Language：English   Publisher：Zoological Society of Japan  

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Other Link： http://id.nii.ac.jp/1141/00038911/

Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (bulletin of university, research institution)  

CiNii Books

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)  

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Language：English   Publisher：Zoological Society of Japan  

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Other Link： http://id.nii.ac.jp/1141/00037945/

Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)  

DOI： 10.11477/mf.2425100728

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Language：Japanese   Publishing type：Article, review, commentary, editorial, etc. (scientific journal)   Publisher：医歯薬出版  

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Other Link： http://search.jamas.or.jp/link/ui/2002083499

Language：Japanese   Publisher：(一社)日本解剖学会  

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Event date： 2016.7

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Here we present the first evidence that muscle-specific kinase (MUSK) antigen can cause myasthenia in animals. MUSK is expressed at the postsynaptic membranes of neuromuscular junctions (NMJ) and forms complexes with acetylcholine receptors (AChR) and rapsyn. MUSK is activated by agrin, which is released from motoneurons, and induces AChR clustering and subsequent formation of NMJ in embryos. Notably, autoantibodies against MUSK were found in a proportion of patients with generalized myasthenia gravis (MG) but without the characteristic AChR autoantibodies. However, MUSK autoantibodies had no known pathogenic potential, and animals immunized with purified MUSK proteins did not develop MG in former studies. In contrast, we have now injected rabbits with MUSK ectodomain protein in vivo and evoked a MG-like muscle weakness with a reduction of AChR clustering at the NMJ. Our results showed that MUSK is required for maintenance of synapses and that interference with that function by MUSK antibodies causes myasthenic weakness. In vitro, AChR clustering in myotubes is induced by agrin and agrin-independent inducers, which do not activate MUSK. Neither the receptor nor the activation mechanisms of AChR clustering induced by agrin-independent inducers has been identified with certainty, but MUSK autoantibodies in myasthenic animals inhibited both agrin and agrin-independent AChR clustering. MUSK plays multiple roles in pre-patterning of the postsynaptic membrane before innervation and formation of NMJ in embryos. Some of these mechanisms may also participate in the maintenance of mature NMJ. This model system would provide new knowledge about the molecular pathogenesis of MG and MUSK functions in mature NMJ.

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