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A novel gene, norbin, increased by treatment of tetraethylammonium in the rat hippocampal slice and accompanied with neurite-outgrowth in Neuro 2a cells
https://ir.soken.ac.jp/records/1099
https://ir.soken.ac.jp/records/1099cf98a25c-75e7-43ae-a9a0-a328746c2b75
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
要旨・審査要旨 / Abstract, Screening Result (364.1 kB)
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| Item type | 学位論文 / Thesis or Dissertation(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2010-02-22 | |||||
| タイトル | ||||||
| タイトル | A novel gene, norbin, increased by treatment of tetraethylammonium in the rat hippocampal slice and accompanied with neurite-outgrowth in Neuro 2a cells | |||||
| タイトル | ||||||
| タイトル | A novel gene, norbin, increased by treatment of tetraethylammonium in the rat hippocampal slice and accompanied with neurite-outgrowth in Neuro 2a cells | |||||
| 言語 | en | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_46ec | |||||
| 資源タイプ | thesis | |||||
| 著者名 |
篠崎, 幸喜
× 篠崎, 幸喜 |
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| フリガナ |
シノザキ, コウキ
× シノザキ, コウキ |
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| 著者 |
SHINOZAKI, Kohki
× SHINOZAKI, Kohki |
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| 学位授与機関 | ||||||
| 学位授与機関名 | 総合研究大学院大学 | |||||
| 学位名 | ||||||
| 学位名 | 博士（学術） | |||||
| 学位記番号 | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | 総研大甲第414号 | |||||
| 研究科 | ||||||
| 値 | 生命科学研究科 | |||||
| 専攻 | ||||||
| 値 | 20 生理科学専攻 | |||||
| 学位授与年月日 | ||||||
| 学位授与年月日 | 1999-03-24 | |||||
| 学位授与年度 | ||||||
| 値 | 1998 | |||||
| 要旨 | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | Selective expression of various genes occurs in the process of development, maintenance, injury, and death of unicellular as well as complex organisms, and proteins encoded by such genes plays an essential role(s) at each steps in such biological activities. Specifically expressed genes are usually identified by differential cloning, such as subtraction cloning. In this study, he has applied subtraction cloning to study the genes, whose expression is specifically accompanied by long - lasting changes in brain functions.<br /> Synaptic plasticity is one of the phenomena directly related to learning and memory in neural pathways. It can be considered to be re - differentiation of neurons in the sense that it is often accompanied with reformation of neuronal circuit. For example, it was reported that synaptic reorganization in hippocampus was induced by kindling, a model of synaptic plasticity, and that sprouting of mossy fiber in hippocampus occurred in long - term potentiation (LTP), an another model of synaptic plasticity. On the other hand, some genes which are related to the formation of neuronal circuit (for example, neurotorophin - 3, heparin - binding growth - associated molecule (HB - GAM), GAP - 43) are known to be related to LTP, in addition to neuronal differentiation. Hence it is considered to be an effective strategy to understand synaptic plasticity to find and analyze the genes related to both synaptic plasticity, such as LTP, and neuronal differentiation.<br /> LTP is a phenomenon representing synaptic plasticity and has been extensively studied as a model of learning and memory. The electric high frequent stimulation of presynaptic cells results in persistent potentiation of response of post synaptic cells (tetanic LTP). Although, during or immediately after the stimulation, the specific gene expression is considered to be activated and to play a role(s) in the maintenance of LTP, little is known about the molecular mechanism. LTP can also be induced by a bath application of tetraethylammonium (TEA), a K<SUP>+</SUP> channel blocker, to the rat hippocampal slices in a similar mechanism to tetanic LTP (TEA - LTP). In contrast to tetanic LTP, which is considered to occur in a limited number of neurons around the electrode, TEA - LTP are considered to involve many neurons. This enables us to find genes related to this phenomenon by the subtraction cloning.<br /> They screened for new genes related to TEA - LTP by subtraction between the mRNA of a rat whole brain and that of the TEA - treated slices. During this screening, a novel gene, named norbin whose mRNA was increased about 1 hr after the TEA - treatment, was identified. It was about 5 kb in length and had no significant homology with known genes. The predicted protein sequence of norbin consisted of 729 amino acids.<br /> Immunoblotting of rat tissues with the antibody against norbin revealed a band of about 80 kDa, which corresponded to the calculate molecular weight of 78,894 Da and was specifically detected in neural tissues. The double staining of the rat brain using anti - GFAP antibody and in situ hybridization of norbin, or anti - CNPase antibody and anti - norbin antibody revealed that norbin was mainly expressed in neurons. Immunostaining of a rat brain using anti - norbin antibody revealed that norbin was distributed throughout the brain. In the cerebral cortex, the strong signal was observed in neurons of the 2<SUP>nd </SUP>- 3<SUP>rd</SUP> layers. In the cerebellum, Purkinje cells and granule cells were stained well. In the hippocampus, pyramidal cells and granule cells were strongly stained. Norbin was localized in the somata except for nuclei and dendrites of neurons. The subcellular fractionation study revealed that norbin is a cytosolic protein.<br /> Transfection of norbin cDNA facilitated process-outgrowth of a neuroblastoma cell line, Neuro 2a cells. In the case of a fibroblast cell line, COS - 1 cells, no effect on their morphology was observed. These results suggest the selective function of norbin in neural tissues.<br /> Immunoblotting of the developing rat showed that faint expression was observed in the whole body on embryonic day (E) 10.5 and in the brain on E12.5. The expression in the brain gradually increased towards the juvenile stage (postnatal day 7 - 28) and decreased thereafter in adult stage, except for the hippocampus, where the expression increased gradually until adult stage. Immunostaining of E14.5 rat revealed that norbin was expressed in neurons in the cortical plate of the telencephalon, but not in the ventricular zone, suggesting that it might be expressed in neurons which migrated from the ventricular zone and undergone terminal differentiation. The double staining of the developing Purkinje cells using anti - calbindin antibody and anti - norbin antibody revealed that it was distributed throughout the dendrites and somata of the cells, in particular, in their growing dendritic trees, suggesting the involvement of norbin in the dendritic outgrowth.<br /> Since norbin was expressed from the early neural developmental stage to adult, localized in the somatodendritic regions of neurons, and showed a neurite-outgrowth-facilitating activity, it might be involved in the neural differentiation and plasticity, by affecting the somatodendritic functions, in particular, contributing to the modification of neural network. | |||||
| 所蔵 | ||||||
| 値 | 有 | |||||
