Studies on the mechanism of signal transduction of Arc, a gene involved in synaptic plasticity

突触可塑性基因Arc信号转导机制研究

• 批准号: 11680646
• 负责人: SUGIURA Hiroko
• 金额: $ 2.37万
• 依托单位: Tokyo Metropolitan Institute for Neurosciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680646/
• 关键词: synaptic plasticity; Arc; dynamin; two-hybrid system; GTPase activity; protein-protein interaction; SH3 protein; endocytosis; SH3タンパク質; two hybrid system

To define the changes in gene expression that may underlie long-lasting behavioral effects by cocaine and methamphetamine, drug-induced immediate early gene Arc has been characterized and proteins are screened the two-hybrid library to identify interacting partners of Arc in our laboratory. I found a novel SH3 protein, SH3P13 bound to Arc in vivo. SH3P13 consists of twoalternative splicing products 13L and 13S, and binds to dynamin that is involved in endocytotic machinery. immunohistochemistry of primary hippocampal neurons showed that both dynamin and SH3P13 proteins were expressed in neurons and co-localized at synapses. Furthermore, immunoelectron microscopy reveal that SH3P13 was localized to postsynaptic spines. It activated the GTPase of dynamin, which is known to regulate dopamine receptor-mediated endocytosis. Therefore, we examined whether overexpression of SH3P13 in COS-7 cells regulates dynamin-dependent internalization of dopamine D2 receptor. Unexpectedly, coexpression of SH3P13 almost completely inhibited sequestration of D2 receptor after dopamine stimulation. These results suggest that SH3P13 negatively regulates dopamine receptor mediated-endocytosis by competing with endogenous dynamin-binding protein(s) in COS-7 cells. Arc protein induced by cocaine or methamphetamine might regulate dynamin-dependent internalization of dopamine receptor via binding to SH3P13. The study of the regulation of SH3P13 on the dopamine-dependent endocytosis by Arc may provide an insight into physiological function of SH3P13 and Arc protein.

为了确定可卡因和甲基苯丙胺造成长期行为影响的基因表达变化，我们对药物诱导的早期基因 Arc 进行了表征，并对双杂交文库中的蛋白质进行了筛选，以鉴定我们实验室中 Arc 的相互作用伙伴。我发现了一种新的 SH3 蛋白，SH3P13 在体内与 Arc 结合。 SH3P13 由两个选择性剪​​接产物 13L 和 13S 组成，并与参与胞吞机制的动力结合。原代海马神经元的免疫组织化学显示，dynamin 和 SH3P13 蛋白均在神经元中表达，并共定位于突触。此外，免疫电子显微镜显示 SH3P13 位于突触后棘。它激活动力的 GTP 酶，已知该酶可调节多巴胺受体介导的内吞作用。因此，我们检查了 COS-7 细胞中 SH3P13 的过度表达是否调节多巴胺 D2 受体的动力依赖性内化。出乎意料的是，SH3P13 的共表达几乎完全抑制了多巴胺刺激后 D2 受体的隔离。这些结果表明，SH3P13 通过与 COS-7 细胞中的内源动力结合蛋白竞争，负向调节多巴胺受体介导的内吞作用。可卡因或甲基苯丙胺诱导的 Arc 蛋白可能通过与 SH3P13 结合来调节多巴胺受体的动力依赖性内化。研究 SH3P13 对 Arc 多巴胺依赖性内吞作用的调节可能有助于深入了解 SH3P13 和 Arc 蛋白的生理功能。

项目成果

Yamagata,K., et al.: "Activation of an effector immediate-early gene arc by methamphetamine,"the Annals of the New York Academy of Science. 914. 22-32 (2000)

Yamagata,K., et al.：“甲基苯丙胺对效应子立即早期基因的激活”，《纽约科学院年鉴》。

Yamagata, K., Suzuki, K., Sugiura, H., Kawashima, N.and Okuyama, S.: "Activation of an effector immediate-early gene arc by methamphetamine."Ann. N Y Acad. Sci.. 914. 22-32 (2000)

Yamagata, K.、Suzuki, K.、Sugiura, H.、Kawashima, N. 和 Okuyama, S.：“甲基苯丙胺激活立即早期基因弧。”Ann。

Yamagata, K., Andreasson, K.I., Sugiura, H., Maru, E., Dominique, M., Irie, Y., Miki, N., Hayashi, Y., Yoshioka, M., Kaneko, K., Kato, H., and Worley, P.F.: "Arcadlin is a neural activity-regulated cadherin involved in long term potentiation."J.Biol. Chem

Yamagata, K.、Andreasson, K.I.、Sugiura, H.、Maru, E.、Dominique, M.、Irie, Y.、Miki, N.、Hayashi, Y.、Yoshioka, M.、Kaneko, K.、Kato

Kanato Yamagata et al: "Slow Synaptic Responses and Modulation"Springer-Verlag. 8 (2000)

Kanato Yamagata 等人：“慢突触反应和调制”Springer-Verlag。

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Yamagata,K., et al.：“内毒素诱导发热期间大鼠脑内皮细胞中微粒体型前列腺素 E 合酶与环氧合酶 2 的共表达。”J.Neuroscience。