2021/11/27 更新

写真a

サトウ ノボル
佐藤 昇
SATO Noboru
所属
教育研究院 医歯学系 医学系列 教授
医学部 医学科 教授
医歯学総合研究科 生体機能調節医学専攻 教授
職名
教授
外部リンク

学位

  • 博士(医学) ( 1993年3月   筑波大学 )

研究分野

  • ライフサイエンス / 解剖学

  • ライフサイエンス / 神経形態学

経歴(researchmap)

  • 福島県立医科大学 医学部 医学科 医学部医学科 神経解剖・発生学講座   准教授

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経歴

  • 新潟大学   医学部 医学科   教授

    2007年4月 - 現在

  • 新潟大学   医歯学総合研究科 生体機能調節医学専攻   教授

    2007年4月 - 現在

学歴

  • 筑波大学   医学研究科   形態系

    - 1993年

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    国名: 日本国

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  • 筑波大学

    - 1993年

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  • 筑波大学

    - 1989年

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  • 筑波大学   医学専門学群

    - 1989年

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    国名: 日本国

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所属学協会

 

論文

  • Development of fin-innervating motor neurons after peripheral target removal in medaka fish 査読

    Akina Chiba, Kenichi Soma, Keisuke Watanabe, Hiroshi Nagashima, Noboru Sato

    Developmental Neurobiology   2020年12月

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    担当区分:責任著者   記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Novel concept for the epaxial/hypaxial boundary based on neuronal development. 査読 国際誌

    Hiroshi Nagashima, Daisuke Koga, Satoshi Kusumi, Katsuki Mukaigasa, Hiroyuki Yaginuma, Tatsuo Ushiki, Noboru Sato

    Journal of anatomy   237 ( 3 )   427 - 438   2020年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Trunk muscles in vertebrates are classified as either dorsal epaxial or ventral hypaxial muscles. Epaxial and hypaxial muscles are defined as muscles innervated by the dorsal and ventral rami of spinal nerves, respectively. Each cluster of spinal motor neurons passing through dorsal rami innervates epaxial muscles, whereas clusters traveling on the ventral rami innervate hypaxial muscles. Herein, we show that some motor neurons exhibiting molecular profiles for epaxial muscles follow a path in the ventral rami. Dorsal deep-shoulder muscles and some body wall muscles are defined as hypaxial due to innervation via the ventral rami, but a part of these ventral rami has the molecular profile of motor neurons that innervate epaxial muscles. Thus, the epaxial and hypaxial boundary cannot be determined simply by the ramification pattern of spinal nerves. We propose that, although muscle innervation occurs via the ventral rami, dorsal deep-shoulder muscles and some body wall muscles represent an intermediate group that lies between epaxial and hypaxial muscles.

    DOI: 10.1111/joa.13219

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  • The fornix acts as a permissive corridor for septal neuron migration beyond the diencephalic-telencephalic boundary. 査読 国際誌

    Keisuke Watanabe, Hirohide Takebayashi, Noboru Sato

    Scientific reports   10 ( 1 )   8315 - 8315   2020年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Neuronal migration is essential for constructing functional neural networks. Two posterior septal (PS) nuclei, the triangular septal nucleus and bed nuclei of the anterior commissure, are involved in fear and anxiety. During development, glutamatergic PS neurons undergo long-distance rostrodorsal migration from the thalamic eminence (TE) of the diencephalon, then settle in the caudalmost telencephalon. However, the developmental behavior of PS neurons and the guidance structures facilitating their migration remain unknown. We previously demonstrated the migration of PS neurons along the fornix, a major efferent pathway from the hippocampal formation. Here, we show that the postcommissural fornix is essential for PS neuron migration which is largely confined to its axonal tract, which grows in the opposite direction as PS neuron migration. Fornical axons reach the TE prior to initiation of PS neuron rostrodorsal migration. Ectopic expression of Semaphorin 3 A in the dorsomedial cortex resulted in defective fornix formation. Furthermore, loss of the postcommissural fornix stalled PS neuron migration resulting in abnormal accumulation near their origin. This suggests that PS neurons utilize the postcommissural fornix as a permissive corridor during migration beyond the diencephalic-telencephalic boundary. This axonal support is essential for the functional organization of the heterogeneous septal nuclear complex.

    DOI: 10.1038/s41598-020-65284-7

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  • Development of a mouse nerve-transfer model for brachial plexus injury. 査読

    Hanako Wakatsuki, Minoru Shibata, Ken Matsuda, Noboru Sato

    Biomedical research (Tokyo, Japan)   40 ( 3 )   115 - 123   2019年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Nerve transfer involves the use of a portion of a healthy nerve to repair an injured nerve, and the process has been used to alleviate traumatic brachial plexus injuries in humans. Study of the neural mechanisms that occur during nerve transfer, however, requires the establishment of reliable experimental models. In this study, we developed an ulnar-musculocutaneous nerve-transfer model wherein the biceps muscle of a mouse was re-innervated using a donor ulnar nerve. Similar muscle action potentials were detected in both the end-to-end suture of the transected nerve (correctrepair) group and the ulnar-musculocutaneous nerve-transfer group. Also, re-innervated acetylcholine receptor (AChR) clusters and muscle spindles were observed in both procedures. There were fewer re-innervated AChR clusters in the nerve transfer group than in the correct repair group at 4 weeks, but the numbers were equal at 24 weeks following surgery. Thus, our ulnar-musculocutaneous nerve-transfer model allowed physiological and morphological evaluation for re-innervation process in mice and revealed the delay of this process during nerve transfer procedure. This model will provide great opportunities to study regeneration, re-innervation, and functional recovery induced via nerve transfer procedures.

    DOI: 10.2220/biomedres.40.115

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  • Diencephalic progenitors contribute to the posterior septum through rostral migration along the hippocampal axonal pathway. 査読 国際誌

    Keisuke Watanabe, Koichiro Irie, Carina Hanashima, Hirohide Takebayashi, Noboru Sato

    Scientific reports   8 ( 1 )   11728 - 11728   2018年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Septal nuclei are telencephalic structures associated with a variety of brain functions as part of the limbic system. The two posterior septal nuclei, the triangular septal nucleus (TS) and the bed nuclei of the anterior commissure (BAC), are involved in fear and anxiety through their projections to the medial habenular nucleus. However, the development of both the TS and BAC remains unclear. Here, we found a novel caudal origin and putative migratory stream of mouse posterior septal neurons arising from the thalamic eminence (TE), a transient developmental structure at the rostral end of the rodent diencephalon. TE-derived cells, which have glutamatergic identity, migrated rostrally and entered the telencephalic territory by passing beneath the third ventricle. Subsequently, they turned dorsally toward the posterior septum. We also observed that TS and BAC neurons in the postnatal septum were labeled with GFP by in utero electroporation into the TE, suggesting a shared origin. Furthermore, TE-derived septal neurons migrated along the fornix, an efferent pathway from the hippocampus. These results demonstrate that posterior septal neurons have a distinct extratelencephalic origin from other septal nuclei. This heterogeneous origin may contribute to neuronal diversity of the septal nuclear complex.

    DOI: 10.1038/s41598-018-30020-9

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  • Motor neurons with limb-innervating character in the cervical spinal cord are sculpted by apoptosis based on the Hox code in chick embryo. 査読 国際誌

    Katsuki Mukaigasa, Chie Sakuma, Tomoaki Okada, Shunsaku Homma, Takako Shimada, Keiji Nishiyama, Noboru Sato, Hiroyuki Yaginuma

    Development (Cambridge, England)   144 ( 24 )   4645 - 4657   2017年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:COMPANY OF BIOLOGISTS LTD  

    In the developing chick embryo, a certain population of motor neurons (MNs) in the non-limb-innervating cervical spinal cord undergoes apoptosis between embryonic days 4 and 5. However, the characteristics of these apoptotic MNs remain undefined. Here, by examining the spatiotemporal profiles of apoptosis and MN subtype marker expression in normal or apoptosis-inhibited chick embryos, we found that this apoptotic population is distinguishable by Foxp1 expression. When apoptosis was inhibited, the Foxp1+ MNs survived and showed characteristics of lateral motor column (LMC) neurons, which are of a limb-innervating subtype, suggesting that cervical Foxp1+ MNs are the rostral continuation of the LMC. Knockdown and misexpression of Foxp1 did not affect apoptosis progression, but revealed the role of Foxp1 in conferring LMC identity on the cervical MNs. Furthermore, ectopic expression of Hox genes that are normally expressed in the brachial region prevented apoptosis, and directed Foxp1+ MNs to LMC neurons at the cervical level. These results indicate that apoptosis in the cervical spinal cord plays a role in sculpting Foxp1+ MNs committed to LMC neurons, depending on the Hox expression pattern.

    DOI: 10.1242/dev.158873

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  • Developmental origin of the clavicle, and its implications for the evolution of the neck and the paired appendages in vertebrates. 査読 国際誌

    Hiroshi Nagashima, Fumiaki Sugahara, Keisuke Watanabe, Masahiro Shibata, Akina Chiba, Noboru Sato

    Journal of anatomy   229 ( 4 )   536 - 48   2016年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    In fish, the pectoral appendage is adjacent to the head, but during vertebrate evolution a long neck region emerged via caudal relocation of the pectoral appendage. The pectoral appendage is comprised of endochondral portions, such as the humerus and the scapula, and a dermal portion, such as the clavicle, that contributes to the shoulder girdle. In the search for clues to the mechanism of the caudal relocation of the pectoral appendage, the cell lineage of the rostral lateral plate mesoderm was analyzed in chickens. It was found that, despite the long neck region in chickens, the origin of the clavicle attached to the head mesoderm ranged between 1 and 14 somite levels. Because the pectoral limb bud and the endochondral pectoral appendage developed on 15-20 and 15-24 somite levels, respectively, the clavicle-forming region corresponds to the embryonic neck, which suggests that the relocation would have been executed by the expansion of the source of the clavicle. The rostral portion of the clavicle-forming region overlaps the source of the cucullaris muscle, embraces the pharyngeal arches caudally, and can be experimentally replaced with the head mesoderm to form the cucullaris muscle, which implies that the mesodermal portion could have been the head mesoderm and that the clavicle would have developed at the head/trunk boundary. The link between the head mesoderm and the presumptive clavicle appears to have been the developmental constraint needed to create the evolutionarily conserved musculoskeletal connectivities characterizing the gnathostome neck. In this sense, the dermal girdle of the ganathostomes would represent the wall of the branchial chamber into which the endochondral pectoral appendage appears to have attached since its appearance in evolution.

    DOI: 10.1111/joa.12502

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  • A specific tripeptidyl substrate for tripeptidyl peptidase activity is effectively hydrolyzed by alanyl aminopeptidase/aminopeptidase N/CD13 in the rat kidney

    Masahiro Shibata, Masato Koike, Satoshi Kusumi, Noboru Sato, Yasuo Uchiyama

    Archives of Histology and Cytology   76 ( 1 )   1 - 8   2016年3月

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    掲載種別:研究論文(学術雑誌)  

    © 2016 by International Society of Histology and Cytology.L-Alanyl-L-alanyl-L-phenylalanine 4-methylcoumaryl-7-amide (AAF-MCA) is one of the classic substrates for use with tripeptidyl peptidases (TPP-I and TPP-II). We have previously clarified the tissue distribution of TPP-I in detail and noted that the protein expression of TPP-I is often incompatible with its enzyme activity. Herein, we describe the unknown peptidase, which could effectively hydrolyze AAF-MCA, in the rat kidney. The peptidase was purified after four chromatography steps, and its enzyme characteristics were elucidated. The peptidase activity was inhibited by amastatin, bestatin, and o-phenanthroline and was also inhibited by zinc and copper ions. The substrate specificity for several monoamino acidic-MCAs revealed that the peptidase had an affinity for alanyl-MCA. The amino terminal amino acid sequence of the peptidase was x-Ala-Pro-x-Leu-Pro-Gly-Ser-Thr-Ser-Ala-Thr-x-x-Ser, where x indicates undetectable amino acid residues, and the antiserum against the peptidase was immunopositive for the brush border of a renal proximal tubule and the small intestine, and the surface membrane of bile canaliculi. These results indicate that the unknown peptidase that hydrolyzed AAF-MCA is the soluble form of aminopeptidase N/CD13, and caution is required when using AAF-MCA as a substrate for tripeptidyl peptidase assays.

    DOI: 10.1679/aohc.76.1

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  • Evidence from cyclostomes for complex regionalization of the ancestral vertebrate brain. 査読 国際誌

    Fumiaki Sugahara, Juan Pascual-Anaya, Yasuhiro Oisi, Shigehiro Kuraku, Shin-ichi Aota, Noritaka Adachi, Wataru Takagi, Tamami Hirai, Noboru Sato, Yasunori Murakami, Shigeru Kuratani

    Nature   531 ( 7592 )   97 - 100   2016年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The vertebrate brain is highly complex, but its evolutionary origin remains elusive. Because of the absence of certain developmental domains generally marked by the expression of regulatory genes, the embryonic brain of the lamprey, a jawless vertebrate, had been regarded as representing a less complex, ancestral state of the vertebrate brain. Specifically, the absence of a Hedgehog- and Nkx2.1-positive domain in the lamprey subpallium was thought to be similar to mouse mutants in which the suppression of Nkx2-1 leads to a loss of the medial ganglionic eminence. Here we show that the brain of the inshore hagfish (Eptatretus burgeri), another cyclostome group, develops domains equivalent to the medial ganglionic eminence and rhombic lip, resembling the gnathostome brain. Moreover, further investigation of lamprey larvae revealed that these domains are also present, ruling out the possibility of convergent evolution between hagfish and gnathostomes. Thus, brain regionalization as seen in crown gnathostomes is not an evolutionary innovation of this group, but dates back to the latest vertebrate ancestor before the divergence of cyclostomes and gnathostomes more than 500 million years ago.

    DOI: 10.1038/nature16518

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  • Endoplasmic Reticulum-Localized Transmembrane Protein Dpy19L1 Is Required for Neurite Outgrowth. 査読 国際誌

    Keisuke Watanabe, Norihisa Bizen, Noboru Sato, Hirohide Takebayashi

    PloS one   11 ( 12 )   e0167985   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    The endoplasmic reticulum (ER), including the nuclear envelope, is a continuous and intricate membrane-bound organelle responsible for various cellular functions. In neurons, the ER network is found in cell bodies, axons, and dendrites. Recent studies indicate the involvement of the ER network in neuronal development, such as neuronal migration and axonal outgrowth. However, the regulation of neural development by ER-localized proteins is not fully understood. We previously reported that the multi-transmembrane protein Dpy19L1 is required for neuronal migration in the developing mouse cerebral cortex. A Dpy19L family member, Dpy19L2, which is a causative gene for human Globozoospermia, is suggested to act as an anchor of the acrosome to the nuclear envelope. In this study, we found that the patterns of exogenous Dpy19L1 were partially coincident with the ER, including the nuclear envelope in COS-7 cells at the level of the light microscope. The reticular distribution of Dpy19L1 was disrupted by microtubule depolymerization that induces retraction of the ER. Furthermore, Dpy19L1 showed a similar distribution pattern with a ER marker protein in embryonic mouse cortical neurons. Finally, we showed that Dpy19L1 knockdown mediated by siRNA resulted in decreased neurite outgrowth in cultured neurons. These results indicate that transmembrane protein Dpy19L1 is localized to the ER membrane and regulates neurite extension during development.

    DOI: 10.1371/journal.pone.0167985

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  • On the homology of the shoulder girdle in turtles. 査読 国際誌

    Hiroshi Nagashima, Fumiaki Sugahara, Masaki Takechi, Noboru Sato, Shigeru Kuratani

    Journal of experimental zoology. Part B, Molecular and developmental evolution   324 ( 3 )   244 - 54   2015年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The shoulder girdle in turtles is encapsulated in the shell and has a triradiate morphology. Due to its unique configuration among amniotes, many theories have been proposed about the skeletal identities of the projections for the past two centuries. Although the dorsal ramus represents the scapular blade, the ventral two rami remain uncertain. In particular, the ventrorostral process has been compared to a clavicle, an acromion, and a procoracoid based on its morphology, its connectivity to the rest of the skeleton and to muscles, as well as with its ossification center, cell lineage, and gene expression. In making these comparisons, the shoulder girdle skeleton of anurans has often been used as a reference. This review traces the history of the debate on the homology of the shoulder girdle in turtles. And based on the integrative aspects of developmental biology, comparative morphology, and paleontology, we suggest acromion and procoracoid identities for the two ventral processes.

    DOI: 10.1002/jez.b.22584

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  • On the homology of the shoulder girdle in turtles. 査読

    Nagashima, H, Sugahara, F, Takechi, M, Sato, N, Kuratani, S

    J. Exp. Zool. B Mol. Dev. Evol.   324 ( 3 )   244 - 254   2015年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The shoulder girdle in turtles is encapsulated in the shell and has a triradiate morphology. Due to its unique configuration among amniotes, many theories have been proposed about the skeletal identities of the projections for the past two centuries. Although the dorsal ramus represents the scapular blade, the ventral two rami remain uncertain. In particular, the ventrorostral process has been compared to a clavicle, an acromion, and a procoracoid based on its morphology, its connectivity to the rest of the skeleton and to muscles, as well as with its ossification center, cell lineage, and gene expression. In making these comparisons, the shoulder girdle skeleton of anurans has often been used as a reference. This review traces the history of the debate on the homology of the shoulder girdle in turtles. And based on the integrative aspects of developmental biology, comparative morphology, and paleontology, we suggest acromion and procoracoid identities for the two ventral processes.

    DOI: 10.1002/jez.b.22584

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  • Comparative study of the shell development of hard- and soft-shelled turtles. 査読 国際誌

    Hiroshi Nagashima, Masahiro Shibata, Mari Taniguchi, Shintaro Ueno, Naoki Kamezaki, Noboru Sato

    Journal of anatomy   225 ( 1 )   60 - 70   2014年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The turtle shell provides a fascinating model for the investigation of the evolutionary modifications of developmental mechanisms. Different conclusions have been put forth for its development, and it is suggested that one of the causes of the disagreement could be the differences in the species of the turtles used - the differences between hard-shelled turtles and soft-shelled turtles. To elucidate the cause of the difference, we compared the turtle shell development in the two groups of turtle. In the dorsal shell development, these two turtle groups shared the gene expression profile that is required for formation, and shared similar spatial organization of the anatomical elements during development. Thus, both turtles formed the dorsal shell through a folding of the lateral body wall, and the Wnt signaling pathway appears to have been involved in the development. The ventral portion of the shell, on the other hand, contains massive dermal bones. Although expression of HNK-1 epitope has suggested that the trunk neural crest contributed to the dermal bones in the hard-shelled turtles, it was not expressed in the initial anlage of the skeletons in either of the types of turtle. Hence, no evidence was found that would support a neural crest origin.

    DOI: 10.1111/joa.12189

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  • Origin of the unique morphology of the shoulder girdle in turtles. 査読 国際誌

    Hiroshi Nagashima, Tatsuya Hirasawa, Fumiaki Sugahara, Masaki Takechi, Ryo Usuda, Noboru Sato, Shigeru Kuratani

    Journal of anatomy   223 ( 6 )   547 - 56   2013年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The shoulder girdle of turtles has a triradiate morphology. Although its dorsal process represents the scapular blade, the skeletal identities of the two ventral processes remain uncertain. To elucidate the question, developmental patterns of the girdles were compared between Chinese soft-shelled turtles, chickens, and mice. Despite the morphological diversity of adults, the initial primordia of the shoulder girdles showed similar morphological patterns. The ventral two processes developed from the anlagen comparable to those of the acromion and the coracoid in other amniotes. The developmental pattern of the acromion is very similar among embryos, whereas that of the coracoid in mammals differs from that in non-mammals, implying that coracoids are not homologous between non-mammals and mammals. Therefore, amniotes have retained the ancestral pattern of the girdle anlage, and the shoulder girdle of turtles has been achieved through a transformation of the pattern in the late ontogenic period.

    DOI: 10.1111/joa.12116

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  • Elucidation of target muscle and detailed development of dorsal motor neurons in chick embryo spinal cord. 査読 国際誌

    Nobumi Kobayashi, Shunsaku Homma, Tomoaki Okada, Tomoyuki Masuda, Noboru Sato, Keiji Nishiyama, Chie Sakuma, Takako Shimada, Hiroyuki Yaginuma

    The Journal of comparative neurology   521 ( 13 )   2987 - 3002   2013年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The avian cervical spinal cord includes motoneurons (MNs) that send their axons through the dorsal roots. They have been called dorsal motoneurons (dMNs) and assumed to correspond to MNs of the accessory nerve that innervate the cucullaris muscle (SAN-MNs). However, their target muscles have not been elucidated to date. The present study sought to determine the targets and the specific combination of transcription factors expressed by dMNs and SAN-MNs and to describe the detailed development of dMNs. Experiments with tracing techniques confirmed that axons of dMNs innervated the cucullaris muscle. Retrogradely labeled dMNs were distributed in the ventral horn of C3 and more caudal segments. In most cases, some dMNs were also observed in the C2 segment. It was also demonstrated that SAN-MNs existed in the ventral horn of the C1-2 segments and the adjacent caudal hindbrain. Both SAN-MNs and dMNs expressed Isl1 but did not express Isl2, MNR2, or Lhx3. Rather, these MNs expressed Phox2b, a marker for branchial motoneurons (brMNs), although the intensity of expression was weaker. Dorsal MNs and SAN-MNs were derived from the Nkx2.2-positive precursor domain and migrated dorsally. Dorsal MNs remain in the ventral domain of the neural tube, unlike brMNs in the brainstem. These results indicate that dMNs and SAN-MNs belong to a common MN population innervating the cucullaris muscle and also suggest that they are similar to brMNs of the brainstem, although there are differences in Phox2b expression and in the final location of each population. J. Comp. Neurol. 521: 2987-3002, 2013. © 2013 Wiley Periodicals, Inc.

    DOI: 10.1002/cne.23326

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  • Neurogenin2 expression together with NeuroM regulates GDNF family neurotrophic factor receptor α1 (GFRα1) expression in the embryonic spinal cord. 査読 国際誌

    Shimada T, Yaginuma H, Sato N, Homma S

    Developmental biology   370 ( 2 )   250 - 63   2012年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1016/j.ydbio.2012.08.002

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  • Bilateral, asymmetric anomalies of the anterior bellies of digastric muscles. 査読

    Yosuke Yamazaki, Masahiro Shibata, Tatsuo Ushiki, Keitaro Isokawa, Noboru Sato

    Journal of oral science   53 ( 4 )   523 - 7   2011年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Bilateral, asymmetric anomalies of the anterior bellies of digastric muscles were observed during dissection of the submental region. Specifically, four extra muscle bundles were found between the anterior bellies of the digastric muscle. Although anomalies of the anterior bellies of digastric muscles are often observed, this complicated pattern of digastric anomalies has not been previously reported. Our findings and previous observations illustrate the morphogenetic complexity of the anterior belly of the digastric muscle derived from the first pharyngeal arch, which gives rise to jaw musculature such as the mylohyoid muscle.

    DOI: 10.2334/josnusd.53.523

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  • Intrasulcal Electrocorticography in Macaque Monkeys with Minimally Invasive Neurosurgical Protocols

    Takeshi Matsuo, Keisuke Kawasaki, Takahiro Osada, Hirohito Sawahata, Takafumi Suzuki, Masahiro Shibata, Naohisa Miyakawa, Kiyoshi Nakahara, Atsuhiko Iijima, Noboru Sato, Kensuke Kawai, Nobuhito Saito, Isao Hasegawa

    Frontiers in Systems Neuroscience   5   2011年

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:Frontiers Media SA  

    DOI: 10.3389/fnsys.2011.00034

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  • Intrasulcal electrocorticography in macaque monkeys 査読

    Keisuke Kawasaki, Takeshi Matsuo, Takahiro Osada, Hirohito Sawahata, Takafumi Suzuki, Masahiro Shibata, Naohisa Miyakawa, Kiyoshi Nakahara, Noboru Sato, Kensuke Kawai, Nobuhito Saito, Isao Hasegawa

    NEUROSCIENCE RESEARCH   71   E413 - E414   2011年

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    記述言語:英語   出版者・発行元:ELSEVIER IRELAND LTD  

    DOI: 10.1016/j.neures.2011.07.1812

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  • ISLR2 expression during the period of naturally occurring cell death in the embryonic nervous system 査読

    Shunsaku Homma, Takako Shimada, Masahiro Shibata, Noboru Sato, Yasuo Uchiyama, Hiroyuki Yaginuma

    NEUROSCIENCE RESEARCH   68   E257 - E257   2010年

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    記述言語:英語   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Conditional RNA interference using a combination of Cre-loxP system and Tol2 transposition is a useful tool for the developmental studies in the chick 査読

    Masahiro Shibata, Kenjiro Ito, Noboru Sato

    NEUROSCIENCE RESEARCH   68   E372 - E372   2010年

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    記述言語:英語   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Efficient gene transfer to developing chick astrocytes by Tol2 transposition 査読

    Masahiro Shibata, Ryosuke Inoue, Noboru Sato

    MECHANISMS OF DEVELOPMENT   126   S319 - S320   2009年8月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

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  • Efficient gene transfer to developing chick astrocytes by Tol2 transposition 査読

    Masahiro Shibata, Ryosuke Inoue, Noboru Sato

    NEUROSCIENCE RESEARCH   65   S88 - S88   2009年

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    記述言語:英語   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Distinct susceptibility of developing neurons to death following Bax overexpression in the chicken embryo

    N Sato, C Sakuma, Y Sato, TW Gould, RW Oppenheim, H Yaginuma

    CELL DEATH AND DIFFERENTIATION   13 ( 3 )   435 - 445   2006年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Bax is a proapoptotic protein that is required for programmed cell death (PCD) of many neuronal populations. Here we show that, during an early period of retinal PCD and in naturally occurring sensory and motor neuron (MN) death in the spinal cord, Bax delivery results in enhanced death of these neural populations. In contrast, Bax overexpression fails to enhance an early phase of MN death that occurs in the cervical spinal cord, although overexpressed Bax appears to be activated in dying MNs. Bax overexpression does not also affect the survival of immature neurons prior to the PCD period. Taken together, these data provide the first in vivo evidence suggesting that Bax appears to act selectively as an executioner only in neurons undergoing PCD. Furthermore, although Bax appears to mediate the execution pathway for PCD, the effect of Bax overexpression on susceptibility to death differs between different neuronal populations.

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  • Early motoneuron death in the cervical spinal cord of the avian embryo occurs in the subgroup of motoneurons that correspond to the motoneurons innervating intercostal muscles 査読

    Hiroyuki Yaginuma, Nobumi Kobayashi, Shunsaku Homma, Noboru Sato, Takako Shimada, Chie Sakuma

    NEUROSCIENCE RESEARCH   55   S120 - S120   2006年

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    記述言語:英語   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Spatial and temporal regulation of gene transfer in the chicken embryo 査読

    Noboru Sato, Chie Sakuma, Hiroyuki Yaginuma

    NEUROSCIENCE RESEARCH   55   S106 - S106   2006年

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  • Gene delivery into the chicken embryo by using replication-competent retroviral vectors. 査読

    Noboru Sato

    Fukushima journal of medical science   50 ( 2 )   37 - 46   2004年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Rous sarcoma virus (RSV)-derived retroviral vectors have allowed for efficient gene transfer into the chicken embryo which is a classical model for studying vertebrate development. Current evidence reveals that this method can be used for regionally restricted expression, inducible expression, and for interfering with endogenous gene function, suggesting that gain-of-function and loss-of-function strategies for specific genes can be achieved spatially and temporally in the avian embryo. Thus, retroviral-mediated gene transfer into the chicken embryo coupled with a wide variety of strategies is now an important tool to address specific biological questions in the vertebrate.

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  • Bcl-2 rescues motoneurons from early cell death in the cervical spinal cord of the chicken embryo. 査読 国際誌

    Noboru Sato, Chie Sakuma, Hiromi Kato, Carolanne E Milligan, Ronald W Oppenheim, Hiroyuki Yaginuma

    Journal of neurobiology   53 ( 3 )   381 - 90   2002年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS INC  

    Motoneurons (MNs) in the cervical spinal cord of the chicken embryo undergo programmed cell death (PCD) between embryonic day (E) 4 and E5. The intracellular molecules regulating this early phase of PCD remain unknown. Here we show that introduction of Bcl-2 by a replication-competent avian retroviral vector prevented MN degeneration at E4.5, whereas the expression of the green fluorescent protein (GFP) was ineffective. Bcl-2 expression did not affect the number of Islet-1/2-positive MNs at the onset of cell death (E4). However, when examined at the end of the cell death period (E5.5), the number of Islet-1/2-positive MNs was clearly increased in Bcl-2-transfected embryos compared with control and GFP-transfected embryos. Activation of caspase-3, which is normally observed in this early MN death, was also prevented by Bcl-2. Thus, MNs in the cervical spinal cord appear to use intracellular pathway(s) for early PCD that is responsive to Bcl-2.

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  • Regulated gene expression in the chicken embryo by using replication-competent retroviral vectors. 査読 国際誌

    Noboru Sato, Kenji Matsuda, Chie Sakuma, Douglas N Foster, Ronald W Oppenheim, Hiroyuki Yaginuma

    Journal of virology   76 ( 4 )   1980 - 5   2002年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER SOC MICROBIOLOGY  

    Rous sarcoma virus (RSV)-derived retroviral vector could efficiently deliver the green fluorescent protein (GFP), which is driven by the internal cytomegalovirus enhancer/promoter, into restricted cell populations in the chicken embryo. RSV-derived vectors coupled with the tet regulatory elements also revealed doxycycline-dependent inducible GFP expression in the chicken embryo in ovo.

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  • Roles of Caspases in the programmed cell death of motoneurons in vivo 査読

    H Yaginuma, N Sato, S Homma, RW Oppenheim

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   64 ( 5 )   461 - 474   2001年12月

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    記述言語:英語   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Cysteine proteases comprising the caspase family have been considered one of the major executioners of programmed cell death. However, detailed analyses of the programmed cell death of developing motoneurons in mice following the genetic deletion of two key caspases, casp-3 and casp-9, and in the chick embryo following treatment with caspase inhibitors, indicate that normal amounts of cell loss occur although the death process is delayed. Motoneurons undergoing programmed cell death without caspase activities exhibit a nonapoptotic morphology in which nuclear changes such as chromatin condensation are absent or reduced and which exhibit extensive cytoplasmic vacuolization such as is rarely observed in degenerating control neurons. These results suggest that caspases are involved in, but are not indispensable for, the developmental death of motoneurons, and that one function of caspases may be to facilitate the removal of cells that are destined to die. Possible alternative caspase-independent pathways for the programmed death of motoneurons are discussed.

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  • Selective localization of Bcl-2 to the inner mitochondrial and smooth endoplasmic reticulum membranes in mammalian cells 査読

    T Gotow, M Shibata, S Kanamori, O Tokuno, Y Ohsawa, N Sato, K Isahara, Y Yayoi, T Watanabe, JF Leterrier, M Linden, E Kominami, Y Uchiyama

    CELL DEATH AND DIFFERENTIATION   7 ( 7 )   666 - 674   2000年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE PUBLISHING GROUP  

    Bcl-2, an anti-apoptotic protein, is believed to be localized in the outer mitochondrial membrane, endoplasmic reticulum, and nuclear envelope, However, Bcl-2 has also been suggested as playing a role in the maintenance of mitochondrial membrane potential, indicating its possible association with the inner mitochondrial membrane. We therefore further examined the exact localization of Bcl-2 in mitochondria purified from wild-type and bcl-2-transfected PC12 cells and pre- and postnatal rat brains. Double immunostaining demonstrated that Bcl-5 was co-localized with subunit beta of F(1)F(0)ATPase in the inner mitochondrial membrane. Biochemical analysis of isolated mitochondria using digitonin and trypsin suggests an association of Bcl-2 with the inner mitochondrial membrane, More interestingly, the majority of Bcl-2 disappeared from the inner membrane of mitochondria when cultured under serum deprivation. These results suggest that Bcl-2 acts as an anti-apoptotic regulator by localizing mainly to the inner mitochondrial and smooth ER membranes.

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  • Novel biphasic effect of pyrrolidine dithiocarbamate on neuronal cell viability is mediated by the differential regulation of intracellular zinc and copper ion levels, NF-kappa B, and MAP kinases 査読

    KC Chung, JH Park, CH Kim, HW Lee, N Sato, Y Uchiyama, YS Ahn

    JOURNAL OF NEUROSCIENCE RESEARCH   59 ( 1 )   117 - 125   2000年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Nuclear factor kappa B (NF-kappa B) is a transcription factor involved in the expression of a wide range of genes, most of which code for proteins that play a role in immunity and inflammation. Pyrrolidine dithiocarbamate (PDTC) is a well-known inhibitor of NF-kappa B. Although its mechanism of action is conferred by its antioxidant property, other mechanisms by which PDTC can act as a prooxidant, metal chelator, and free thiol group modulator have recently been suggested. Here we report that PDTC caused a dual effect on cell viability in neuronal rat pheochromocytoma (PC12) cells, depending on its concentration. Increase of intracellular zinc and copper ion levels selectively potentiated the cytotoxic PDTC effect in a dose-dependent manner, and thiol reagents, such as glutathione and N-acetylcysteine, as well as divalent metal-chelating reagents, such as EDTA and bathocuproline disulfonic acid, blocked its cell death effect. The differential effect of PDTC on cell viability correlates well with the inhibition of NF-kappa B activities. In addition, PDTC differentially activated microtubule-associated protein (MAP) kinases, such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), but not p38, depending on its dose, and the coaddition of glutathione (GSH), other antioxidants, and metal ions also modulated their activities. Furthermore, stable Bcl-2 expression blocked the PDTC-induced cell death. These results suggest that the thiol groups and free zinc and copper ion levels are important for the novel biphasic PDTC effect on cell viability, which is associated with the differential activation of NF-kappa B and MAP kinases. (C) 2000 Wiley-Liss, Inc.

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  • Regulation of a novel pathway for cell death by lysosomal aspartic and cysteine proteinases 査読

    K Isahara, Y Ohsawa, S Kanamori, M Shibata, S Waguri, N Sato, T Gotow, T Watanabe, T Momoi, K Urase, E Kominami, Y Uchiyama

    NEUROSCIENCE   91 ( 1 )   233 - 249   1999年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    PC12 cells undergo apoptosis when cultured under conditions of serum deprivation. In this situation, the activity of caspase-3-like proteinases was elevated, and the survival rate could be maintained by treatment with acetyl-DEVD-cho, a specific inhibitor of caspase-3. In a culture of PC12 cells treated with acetyl-DEVD-cho, where caspase-3-like proteinases are not activated, CA074, a specific inhibitor of cathepsin B induced active death of the cells. Cathepsin B antisense oligonucleotides showed a similar effect to CA074 on the induction of active cell death. By double staining of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling and activated caspase-3, the dying cells treated with CA074 were positive for terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling staining but negative for activated caspase-3. Ultrastructurally, the cells were relatively large and had nuclei with chromatin condensation. The initiation of cell death by CA074 or the cathepsin B antisense were inhibited by the addition of pepstatin A, a lysosomal aspartic proteinase inhibitor, or by cathepsin D antisense. To examine whether this cell death pathway was present in cell types other than PC12 cells, we analysed dorsal root ganglion neurons obtained from rat embryos on the 15th gestational day, a time when they require nerve growth factor for survival and differentiation in culture. When cultured in the absence of nerve growth factor, the neurons survived in the presence of acetyl-DEVD-cho or acetyl-YVAD-cho. Under these conditions, CA074 reduced the survival rate of the neurons, which was subsequently restored by the further addition of pepstain A.
    These results suggest that a novel pathway for initiating cell death exists which is regulated by lysosomal cathepsins, and in which cathepsin D acts as a death factor. We speculate that this death-inducing activity is normally suppressed by cathepsin B. (C) 1999 IBRO. Published by Elsevier Science Ltd.

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  • A novel strategy for introducing exogenous Bcl-2 into neuronal cells: The Cre/loxP system-mediated activation of Bcl-2 for preventing programmed cell death using recombinant adenoviruses

    N Sato, SW Wang, L Li, K Okabe, M Hashimoto, H Yaginuma, K Mikoshiba, Y Uchiyama, T Uetsuki, K Yoshikawa, CE Milligan, RW Oppenheim

    MOLECULAR AND CELLULAR NEUROSCIENCE   12 ( 1-2 )   65 - 78   1998年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC  

    We have established a novel strategy for introducing exogenous Bcl-2 into neuronal cells that is mediated by Cre/loxP recombination using recombinant adenoviral vectors. An on/off-switching cassette for Bcl-2 (CALNLbcl-2) was designed to express Bcl-2 by recombinase Cre-mediated excisional deletion of a spacer DNA flanked by a pair of loxP sites. Exogenous Bcl-2 was clearly induced in PC12 cell lines carrying CALNLbcl-2 after infection with recombinant adenovirus producing recombinase Cre (AxCANCre). Dual infection with both AxCANCre and a recombinant adenovirus bearing CALNLbcl-2 showed efficient delivery of exogenous Bcl-2 into a hybrid motoneuronal cell line and primary chicken spinal motoneurons. The delivery of foreign Bcl-2 promoted survival of motoneurons in medium either containing or lacking trophic support. Thus, this strategy for delivery of exogenous Bcl-2 will be useful for studying neuronal death as well as for introducing foreign genes into postmitotic neurons under the control of recombinase Cre.

    DOI: 10.1006/mcne.1998.0703

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  • Suppression of lysosomal proteolysis at three different steps in regenerating rat liver 査読

    Watanabe K, Ishidoh K, Ueno T, Sato N, Kominami E

    Journal of Biochemistry   124 ( 5 )   947 - 956   1998年

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  • Cloning and expression of the cDNA encoding rat caspase-2

    Noboru Sato, Carolanne E. Milligan, Yasuo Uchiyama, Ronald W. Oppenheim

    Gene   202 ( 1-2 )   127 - 132   1997年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We have isolated cDNA clones for rat caspase-2 (also called Nedd2/Ich-1), that encodes a protein similar to interleukin-1β-converting enzyme (ICE) and the product of the nematode Caenorhabditis elegans cell death gene ced-3 both of which play an important role in programmed cell death (PCD). The rat caspase-2 cDNA clones have an open reading frame (ORF) of 452 amino acids (aa). The predicted aa sequence of rat caspase-2 is highly similar to that of mouse Nedd2 (97.3%) and human Ich-1(L) (91.3%). The aa sequence QACRG containing the active Cys residue, that is necessary for the proteolytic activity of ICE/Ced-3 (caspase) family of proteases, is also conserved in rat caspase-2. Rat caspase-2 also has several Asp residues in the amino and carboxyl cleavage regions similar to other caspase family proteins. We have developed PC12 cells carrying an on/off switching cassette of caspase-2 (named PC-Nd cells), which contains the neo gene flanked by a pair of loxP sites, the Cre-specific recognition sequence of 34 nucleotides (nt), that lies between the promoter and the caspase-2 cDNA. This expression cassette was designed to express the neo gene initially and to turn on the expression of caspase-2 by site-specific recombinase Cre-mediated excisional deletion of the neo gene. After infection with Cre-producing recombinant adenovirus (re-Ad), the expression of caspase-2 was highly induced in PC-Nd cells and presumptive actively processed fragments of caspase-2 were also observed. This gene activation strategy of caspase-2 will be useful for the study of the biological effects of caspase family proteins in PCD.

    DOI: 10.1016/S0378-1119(97)00463-0

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  • Lysosomal cysteine and aspartic proteinases and ubiquitin in rat and human urinary bladder epithelium 査読

    H Tokunaga, S Waguri, N Sato, Y Ohsawa, Y Banya, E Kominami, Y Uchiyama

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   59 ( 3 )   249 - 260   1996年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    To examine localization of cysteine and aspartic proteinases, and ubiquitin in rat and human urinary bladders, immunocytochemistry was applied to the tissues, In semi-thin sections, immunoreactivity for cathepsins B and D was densely localized throughout epithelial layers of rats and humans, while that for cathepsins H and L was mainly localized in rat superficial and human intermediate cells, Immunoreactivity for cathepsin C was relatively high in rat and human epithelia, especially in humans, Immunoreactivity for ubiquitin was detected in rat and human epithelial cells. By electron microscopy, vesicular or heterogeneously dense lysosomes labeled with immunogold particles indicating cathepsin B were seen in rat and human epithelial cells; particularly, they often appeared near fusiform vesicles in rat superficial cells and in human intermediate and superficial cells. By double immunostaining, lysosomes with or without vesicular structures were co-labeled with immunogold particles showing both cathepsin B and ubiquitin, The results suggest that cathepsins B, C, H, and L, and cathepsin D are involved in the lysosomal system of rat and human bladder epithelia, Moreover, considering that ubiquitin is a cofactor in the soluble ATP-dependent proteolysis, the results may also indicate that epithelial cells actively form autophagolysosomes.

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  • Participation of cathepsins B, H, and L in perikaryal condensation of Ca1 pyramidal neurons undergoing apoptosis after brief ischemia 査読

    T Nitatori, N Sato, E Kominami, Y Uchiyama

    INTRACELLULAR PROTEIN CATABOLISM   389   177 - 185   1996年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:PLENUM PRESS DIV PLENUM PUBLISHING CORP  

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  • CYSTEINE PROTEINASES IN GH(4)C(1) CELLS, A RAT PITUITARY-TUMOR CELL-LINE, ARE SECRETED BY THE CONSTITUTIVE AND REGULATED SECRETORY PATHWAYS 査読

    S WAGURI, N SATO, T WATANABE, K ISHIDOH, E KOMINAMI, K SATO, Y UCHIYAMA

    EUROPEAN JOURNAL OF CELL BIOLOGY   67 ( 4 )   308 - 318   1995年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WISSENSCHAFTLICHE VERLAG GMBH  

    Secretory granules of GH(4)C(1) cells, a rat pituitary tumor cell line, are known to be induced by the treatment of estradiol (E(2)), insulin, and epidermal growth factor (EGF). We examined changes in the localization of cathepsins B, H, and L, lysosomal cysteine proteinases, in GH(4)C(1) cells before and after hormonal treatment. Northern blotting and immunofluorescence microscopy showed that both mRNAs and intracellular protein concentrations of these enzymes were increased in the hormone-induced cells, By immunoelectron microscopy, immunogold particles indicating cathepsins B, H, and L were localized not only in lysosomes but also in some secretory granules. To further examine the molecular forms of these proteinases in secretory granules, radiolabeling and immunoprecipitation methods were applied to the media of the cells incubated with or without secretagogues (100 nM 12-O-tetradecanoylphorbol-13-acetate and 50 mu M forskolin); the proforms of cathepsins B, H, and L were secreted from the cells by the constitutive pathway, whereas the mature forms of cathepsins B and H, and the preform and mature form of cathepsin L were secreted by the regulated pathway. These results suggest that in hormone-induced GH(4)C(1) cells, cathepsins B, H, and L are sorted from the Golgi complex not only into lysosomes but also into secretory granules, in which proforms of cathepsins B and H, and a part of procathepsin L are processed into mature forms.

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  • THE FATE OF EFFETE EPITHELIAL-CELLS AT THE VILLUS TIPS OF THE HUMAN SMALL-INTESTINE 査読

    T SHIBAHARA, N SATO, S WAGURI, T IWANAGA, A NAKAHARA, H FUKUTOMI, Y UCHIYAMA

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   58 ( 2 )   205 - 219   1995年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Until recently, little has been known about the morphological features of dying enterocytes at the villus tips of the human small intestine. The present study aimed to show the exfoliating processes of effete enterocytes at the villus tips. Cellular elements of the duodenal lumen and jejunal tissue in humans were fixed and processed for DNA nick end labeling (TUNEL), and transmission and scanning electron microscopy (TEM and SEM). Most cellular elements in the duodenal lumen were enterocytes having TUNEL-positive nuclei. By SEM, protruding enterocytes were discerned at the villus tips. Using the SEM samples embedded in epoxy resin, protruding enterocytes were observed at the villus tips by TEM; they were shrunk by forming numerous clear and autophagic vacuoles, took dome-like profiles, and possessed nuclei with chromatin condensation. The intercellular spaces beneath these protruding or effete enterocytes were often occupied by large lymphocytes. By TUNEL reaction, positive stainings appeared in the epithelium not only at the tip of the villi but also around the site. The results suggest that effete enterocytes at the villus tips of human small intertine are first shrunk by forming clear and autophagic vacuoles, and showed that their nuclei exhibit chromatin condensation immediately before being exfoliated into the lumen.

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  • APOPTOSIS AND HETEROPHAGY OF MEDIAL EDGE EPITHELIAL-CELLS OF THE SECONDARY PALATINE SHELVES DURING FUSION 査読

    K TANIGUCHI, N SATO, Y UCHIYAMA

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   58 ( 2 )   191 - 203   1995年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Recent in vivo and in vitro studies have suggested that medial edge epithelial (MEE) cells covering the lateral palatine shelves do not undergo cell death, but migrate into the oral and nasal epithelium or transform into mesenchymal cells. We, therefore, reexamined the fate of MEE cells during palatal fusion in rat embryos by in situ 3' nick end labeling of dUTP (TUNEL), electron microscopy, and immunohisto/cytochemistry. TUNEL staining revealed positive nuclei in the medial edge epithelium immediately prior to contact, in epithelial triangles formed between the epithelial seam and nasal or oral epithelium, in epithelial pearls, and in mesenchymal tissue near the epithelium. However, these TUNEL-positive cells were rarely present in the epithelial seam. Electron microscopy revealed MEE cells showing nuclear chromatin condensation and cell shrinkage, and apoptotic bodies in the fusing epithelium; these often contained apoptotic body-like structures as heterophagosomes. By double staining using a laser scanning microscope, TUNEL-positive nuclei mere co-localized with lysosomal cysteine proteinases, cathepsin B or L in MEE and mesenchymal cells adjacent to the epithelium. These results suggest that MEE cells undergo apoptosis during the palatal formation, even though they migrate into epithelial triangles or transform into mesenchymal cells. Moreover, apoptotic bodies and cellular debris were phagocytosed by adjacent MEE cells or mesenchymal cells and digested by lysosomal enzymes.

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  • DELAYED NEURONAL DEATH IN THE CA1 PYRAMIDAL CELL LAYER OF THE GERBIL HIPPOCAMPUS FOLLOWING TRANSIENT ISCHEMIA IS APOPTOSIS 査読

    T NITATORI, N SATO, S WAGURI, Y KARASAWA, H ARAKI, K SHIBANAI, E KOMINAMI, Y UCHIYAMA

    JOURNAL OF NEUROSCIENCE   15 ( 2 )   1001 - 1011   1995年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS INC  

    The CA1 pyramidal neurons in the hippocampus are selectively vulnerable to transient ischemic damage. in experimental animals, the CA1 pyramidal neurons undergo cell death several days after brief forebrain ischemia. It remains, however, unknown whether this delayed neuronal death is necrosis or apoptosis. To investigate the degenerating processes of the CA1 pyramidal neurons in gerbil hippocampus after brief ischemia, lysosomal and nuclear alterations in the cells were examined using immunocytochemistry, in situ nick-end labeling, and Southern blotting. By light and electron microscopy, immunoreactivity for cathepsins B, H, and L, representative lysosomal cysteine proteinases, increased in the CA1 pyramidal neurons 3 d after ischemic insult, which showed cell shrinkage. By morphometric analysis, the volume density of cathepsin B-positive lysosomes markedly increased 3 d after ischemic insult, while that of autophagic vacuole-like structures also increased at this stage, suggesting that cathepsin B-immunopositive lysosomes increasing in the neurons after ischemic insult are mostly autolysosomes. Nuclei of the CA1 neurons were nick-end labeled by biotinylated dUTP mediated by terminal deoxy-transferase 3 and 4 d after ischemic insult, but not in the prior stages. Simultaneously, dense chromatin masses appeared in nuclei of the neurons. By Southern blotting, laddering of DNA occurred only in CA1 hippocampal tissues obtained 4 d after ischemic insult. Confocal laser scanning microscopy demonstrated that the fragmented DNA in the CA1 pyramidal layer was phagocytosed by microglial cells. The results suggest that delayed death of the CA1 pyramidal neurons after brief ischemia is not necrotic but apoptotic.

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  • NEURONAL DIFFERENTIATION OF PC12 CELLS AS A RESULT OF PREVENTION OF CELL-DEATH BY BCL-2

    N SATO, K HOTTA, S WAGURI, T NITATORI, K TOHYAMA, Y TSUJIMOTO, Y UCHIYAMA

    JOURNAL OF NEUROBIOLOGY   25 ( 10 )   1227 - 1234   1994年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:JOHN WILEY & SONS INC  

    Rat pheochromocytoma PC12 cells die when cultured in serum-free medium. Neurotrophic factors can rescue PC12 cells from cell death, and induce neuronal differentiation. To further investigate the relationship among cell death, survival, and differentiation, the bcl-2 cDNA, which is known to prevent apoptosis in various types of cells, was transfected into PC12 cells. Six monoclonal bcl-2-transfected cell lines were isolated and confirmed to express mRNA and protein product of bcl-2. The wildtype and bcl-2-transfected PC12 cells were kept to adhere to collagen-coated dishes at the inintiation of serum-free experiments to avoid cellular damage due to detachment of the cells by trituration. Even under the conditions, the control PC12 cells mostly died within 24 h, when cultured in serum-free medium, whereas those expressing Bcl-2 survived even for 7 days in serum-free medium. Moreover, outgrowth of long processes in the bcl-2-transfected cells was only observed under the condition to keep the cells attached to the dishes in serum-free medium without any additive neurotrophic or growth factors. Neurofilament medium protein, which is a neuron-specific cytoskeletal component, was also expressed in the differentiated cells, suggesting that the long processes in bcl-2-transfected PC12 cells are neurites. However, neuronal differentiation of PC12 cells expressing Bcl-2 was not observed when cultured in serum-containing medium. Accordingly, survival of PC12 cells expressing Bcl-2 under the condition which cells usually die may be accompanied with neuronal differentiation. (C) 1994 John Wiley and Sons, Inc.

    DOI: 10.1002/neu.480251005

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  • Cell and tissue distribution of lysosomal cysteine proteinases, cathepsins B, H, and L, and their biological roles 査読

    Uchiyama Y, Waguri S, Sato N, Watanabe T, Ishido K, Kominami E

    Acta Histochemica et Cytochemica   27 ( 4 )   287 - 308   1994年

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    記述言語:英語  

    DOI: 10.1267/ahc.27.287

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  • COEXISTENCE OF RENIN AND CATHEPSIN-B IN SECRETORY GRANULES OF GRANULAR DUCT CELLS IN MALE-MOUSE SUBMANDIBULAR-GLAND 査読

    K SANO, S WAGURI, N SATO, E KOMINAMI, Y UCHIYAMA

    JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY   41 ( 3 )   433 - 438   1993年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:HISTOCHEMICAL SOC INC  

    Cathepsin B, a representative lysosomal cysteine proteinase, has been demonstrated to coexist with renin in secretary granules of rat pituitary LH/FSH cells and renal juxtaglomerular cells. We investigated immunocytochemically the localization of cathepsins B, H, and L in the submandibular gland of male mice, in which active renin is also produced. By light microscopy, granular immunodeposits for cathepsin B were detected in epithelial cells of the gland, particularly in granular duct cells and interstitial cells. Immunoreactivity for cathepsins H and L was mainly found in interstitial cells, although that for cathepsin H was weakly seen in acinar cells. By electron microscopy, immunogold particles indicating cathepsin B intensely labeled small granules near the Golgi complex of granular duct cells and weakly labeled large secretory granules, whereas those showing renin labeled both granules. Double immunostaining co-localized immunogold particles showing renin and cathepsin B in small perinuclear granules near the Golgi complex. Some immunopositive granules seemed to be closely associated with the Golgi elements. These results indicate that the co-localization of renin and cathepsin B is also seen in secretory granules of granular duct cells in the mouse submandibular gland, as seen in rat juxtaglomerular and LH/FSH cells. This suggests that cathepsin B is one of the possible candidates for the renin-processing enzyme.

    DOI: 10.1177/41.3.8429206

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  • STRUCTURAL ORGANIZATION OF THE GENE ENCODING RAT CYSTATIN-BETA

    N SATO, K ISHIDOH, Y UCHIYAMA, E KOMINAMI

    GENE   114 ( 2 )   257 - 260   1992年5月

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    記述言語:英語   出版者・発行元:ELSEVIER SCIENCE BV  

    A genomic DNA clone encompassing the gene (cy-beta) encoding rat cystatin-beta (Cy-beta) was isolated by screening with a rat cy-beta cDNA as a probe. The gene spans about 2.6 kb and comprises three exons. The first intron is located between Lys22 and Val23 and the second between LyS56 and Val57 in the deduced amino acid sequence of Cy-beta. The second exon contains the highly conserved QVVAG sequence which, unlike the sequence of other cystatin family members, is not split by an intron. In the 5&apos;-upstream region, three SP-1-binding sites exist, but no typical TATA-box or CAAT-box sequences are found. The difference in the organization of the rat cy-beta gene, encoding a family-1 cystatin, from that encoding members of the other cystatin families, suggests that cy-beta diverged from a common ancestral gene earlier than the separation of genes encoding family-2 and family-3 cystatins.

    DOI: 10.1016/0378-1119(92)90584-C

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  • IMMUNOCYTOCHEMICAL LOCALIZATION OF CATHEPSIN-B IN RAT ANTERIOR-PITUITARY ENDOCRINE-CELLS, WITH SPECIAL REFERENCE TO ITS COLOCALIZATION WITH RENIN AND PRORENIN IN GONADOTROPHS 査読

    Y UCHIYAMA, M NAKAJIMA, T WATANABE, S WAGURI, N SATO, M YAMAMOTO, Y HASHIZUME, E KOMINAMI

    JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY   39 ( 9 )   1199 - 1205   1991年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:HISTOCHEMICAL SOC INC  

    We examined by immunocytochemistry the localization of cathepsin B in endocrine cells of rat anterior pituitary lobe, using a monospecific antibody to cathepsin B. By light microscopy, granular immunodeposits for cathepsin B were detected in most endocrine cells of anterior pituitary lobe. Cells immunoreactive for luteinizing hormone (LH) were diffusely immunostained by anti-cathepsin B. By electron microscopy, immunogold particles for cathepsin B were localized in lysosomes of thyrotrophs, somatotrophs, and mammotrophs. In mammotrophs, immunogold particles for cathepsin B were also detected in crinophagic bodies. Double immunostaining co-localized immunogold particles for LH and cathepsin B in secretory granules of gonadotrophs. Immunocytochemistry was also applied to demonstrate localization of renin and prorenin in LH-producing gonadotrophs; immunogold particles for renin were co-localized with those for LH, cathepsin B, or prorenin in their secretory granules. Immunogold particles for prorenin were also co-localized with those for LH or cathepsin B in secretory granules, but prorenin-positive granules appeared less frequently than renin-positive granules. These results suggest that cathepsin B not only plays a role in the protein degradation in lysosomes of anterior pituitary endocrine cells but also participates in the activation of renin in gonadotrophs, as has been demonstrated in secretory granules of juxtaglomerular cells.

    DOI: 10.1177/39.9.1918937

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  • CYSTEINE PROTEINASES IN BRONCHOALVEOLAR EPITHELIAL-CELLS AND LAVAGE FLUID OF RAT LUNG 査読

    Y ISHII, Y HASHIZUME, T WATANABE, S WAGURI, N SATO, M YAMAMOTO, S HASEGAWA, E KOMINAMI, Y UCHIYAMA

    JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY   39 ( 4 )   461 - 468   1991年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:HISTOCHEMICAL SOC INC  

    We examined the presence of cathepsins B, H, and L in bronchoalveolar epithelial cells, including alveolar macrophages, and in bronchoalveolar lavage fluid (BALF), using immunocytochemistry and immunoblotting. By light and electron microscopy, immunoreactivity for cathepsins B, H, and L was detected in lysosomes of ciliated and non-ciliated epithelial cells of bronchi and bronchioles, and in macrophages. Immunodeposits for cathepsin H only were demonstrated in lamellar bodies of Type II alveolar epithelial cells, suggesting the cosecretion of surfactants and cathepsin H from the cells into the alveolar space. By immunoblotting, cathepsins B and H were found to be present in BALF. To further investigate the origin of these enzymes in BALF, alveolar macrophages obtained from BALF were cultured for 6 hr in a serum-free medium. Immunoblotting revealed that protein bands corresponding to the pro-form and mature form of cathepsin B and the mature form of cathepsin H were present in the culture medium. From these results, the presence of cathepsins B and H in BALF can be explained by the fact that cathepsin B is secreted from alveolar macrophages and cathepsin H is secreted mainly with surfactants from Type II cells and also from macrophages.

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  • Molecular cloning of cDNA for rat cathepsin C: Cathepsin C, A cysteine proteinase with an extremely long propeptide 査読

    Ishidoh K, Muno D, Sato N, Kominami E

    Journal of Biological Chemistry   266 ( 25 )   16312 - 16317   1991年

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  • MOLECULAR-CLONING AND SEQUENCING OF CDNA FOR RAT CYSTATIN-BETA

    N SATO, K ISHIDOH, Y UCHIYAMA, E KOMINAMI

    NUCLEIC ACIDS RESEARCH   18 ( 22 )   6698 - 6698   1990年11月

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    記述言語:英語   出版者・発行元:OXFORD UNIV PRESS UNITED KINGDOM  

    DOI: 10.1093/nar/18.22.6698

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  • IMMUNOCYTOCHEMICAL LOCALIZATION OF CATHEPSIN-B AND CATHEPSIN-H IN CORTICOTROPHS AND MELANOTROPHS OF RAT PITUITARY-GLAND 査読

    Y UCHIYAMA, M NAKAJIMA, D MUNO, T WATANABE, Y ISHII, S WAGURI, N SATO, E KOMINAMI

    JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY   38 ( 5 )   633 - 639   1990年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:HISTOCHEMICAL SOC INC  

    DOI: 10.1177/38.5.2159033

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書籍等出版物

  • 培養細胞への遺伝子導入

    学際企画・組織細胞化学2000  2000年 

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MISC

  • Regulated gene expression in the chicken embryo by using replication-competent retroviral vectors

    N Sato, K Matsuda, C Sakuma, DN Foster, RW Oppenheim, H Yaginuma

    JOURNAL OF VIROLOGY   76 ( 4 )   1980 - 1985   2002年2月

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    記述言語:英語   出版者・発行元:AMER SOC MICROBIOLOGY  

    Rous sarcoma virus (RSV)-derived retroviral vector could efficiently deliver the green fluorescent protein (GFP), which is driven by the internal cytomegalovirus enhancer/promoter, into restricted cell populations in the chicken embryo. RSV-derived vectors coupled with the tet regulatory elements also revealed doxycycline-dependent inducible GFP expression in the chicken embryo in ovo.

    DOI: 10.1128/JVI.76.4.1980-1985.2002

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  • Bcl-2の抗アポトーシス作用発現とミトコンドリア内膜への局在

    金森 市朗, 後藤 隆洋, 柴田 昌宏, 井佐原 京子, 大沢 良之, 佐藤 昇, 渡部 剛, 内山 安男

    神経化学   37 ( 3 )   370 - 370   1998年9月

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    記述言語:日本語   出版者・発行元:日本神経化学会  

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講演・口頭発表等

  • 神経系形成におけるRhoシグナル伝達の重要性

    第112回日本解剖学会総会・全国学術集会  2007年 

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    会議種別:ポスター発表  

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  • Distinct susceptibility of developing neurons to death after Bax overexpression in the chicken embryo

    36th Annual Meeting,Society for neuroscience  2006年 

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    会議種別:ポスター発表  

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  • Spatial and temporal regulation of gene transfer in the chicken embryo.

    2006年 

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    会議種別:ポスター発表  

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  • Distinct susceptibility of developing neurons to death after Bax overexpression in the chicken embryo

    36th Annual Meeting,Society for neuroscience  2006年 

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    会議種別:ポスター発表  

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  • ニワトリ胚での空間的・時間的な遺伝子発現の制御

    日本解剖学会  2006年 

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  • Spatial and temporal regulation of gene transfer in the chicken embryo.

    2006年 

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    会議種別:ポスター発表  

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共同研究・競争的資金等の研究

  • ウイルスベクタ-による遺伝子導入

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    資金種別:競争的資金

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  • Gene transfer by viral vectors

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    資金種別:競争的資金

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  • Neuronal cell death

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    資金種別:競争的資金

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  • 神経細胞死

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    資金種別:競争的資金

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