| Co-Investigator(Kenkyū-buntansha) |
SAKANAKA Masahiro Ehime University Faculty of Medicine, Professor, 医学部, 教授 (60170601)
NISHIMURA Hiroyuki Hyogo College of Medicine, instructor, 医学部, 助手 (20248131)
MATSUYAMA Tomohiro Hyogo College of Medicine, Assistant Professor, 医学部, 講師 (10219529)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Research Abstract |
We localized Mint isoforms in mouse brain. By in situ hybridization, mRNA encoding Mint1 or Mint2 was expressed in neurons throughout the brain. Mint1 mRNA expression was greatest in the limbic system including cingulate cortex, hippocampus, anterior thalamic nuclei, medial habenular nucleus, and mammillary body. Mint2 mRNA was rich in cerebral cortex, entorhinal cortex, and hippocampus, but less prominent in other limbic structures. Mint1 and Mint2 mRNA was distributed among hippocampal pyramidal neurons, while Mint2 mRNA was especially abundant in CA3. Mint1, but not Mint2 mRNA, was abundant in the substantia nigra pars compacta. Immunohistochemistry visualized Mint proteins in axon terminals and neuronal somata, generally following mRNA distribution. In the hippocampus, Mint1 was rich in the entorhinal projections and mossy fibers of the dentate gyrus, while Mint2 was rich in commisural fibers from the contralateral hippocampus and in CA1. Mint1 intensely stained catecholamine-conta
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ining neurons such as the substantia nigra pars compacta, ventral tegmental area, ans locus ceruleus. Central nervous system neurons, then, predominantly express either Mint1 or Mint2. Mints may be involved in transport and localization of neuronal membrane proteins in specific sites of nerve terminals, thereby regulating signals through synapses. We examined the detailed expression pattern of EHSH1/Intersectin, a novel neuronal adaptor protein linking clathrin-mediated endocytosis and signaling pathways, such as the MAPK pathway, receptor-tyrosine kinase/ras-mediated pathway, and rho family of the GTPase-dependent pathway. Separate cultures of neurons, astrocytes, and microglia revealed that the long isoform EHSH1-1 is a primary isoform in the adult CNS and that EHSH1-1 is highly enriched in neurons. Immunohistochemistry of a series of brain sections showed widespread distribution of EHSH1 throughout the brain, with being particularly enriched in Layer III of the neocortex, hippocampus, globus pallidus, subthalamic nucleus, and substantia nigra, which may reflect various conditions in clathrin-mediated endocytosis and the following signal transduction. Less
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