AMPA receptor and its role in astroglia

AMPA受体及其在星形胶质细胞中的作用

• 批准号: 11680815
• 负责人: SEKIGUCHI Masayuki
• 金额: $ 2.24万
• 依托单位: National Center of Neurology and Psychiatry
• 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-11680815/
• 关键词: Astroglia; AMPA receptor; PEPA; Cyclothiaizde; GDNF; Cyclthiazide; 脱感作; カルシウム

We have prepared astroglia-rich cultures from the embryonic rat cerebral cortex, and investigated activation of AMPA receptor in astroglia using calcium-imaging. In our culture condition, AMPA receptor-evoked increases in intracellular calcium concentration was observed in a small population of cells. This calcium responses were potentiated by PEPA and cyclothiazide, allosteric potentiators of AMPA receptors, and the extent of potentiation by two drugs depends upon cells. This is probably due to cell-dependent heterogeneity of AMPA receptors. In these cells, membrane currents mediated by AMPA receptors were recorded using patch clamp recordings.The current responses were also potentiated by PEPA, and show inwardly rectifying I-V relation, at least in the cells we tested. We have examined gene expression stimulated by the activation of glial AMPA receptors. For this purpose, quantitative RT-PCR was carried out for mRNA from glial cultures. We could not detect significant increases in expression of GDNF mRNA level in our cultures. We have further investigated on other gene, and prepared glial cells from other brain regions, in order to get further information on glial AMPA receptors.

我们从胚胎大鼠大脑皮层制备了富含星形胶质细胞的培养物，并使用钙成像研究了星形胶质细胞中 AMPA 受体的激活。在我们的培养条件下，在一小群细胞中观察到 AMPA 受体引起的细胞内钙浓度增加。这种钙反应被 PEPA 和环噻嗪（AMPA 受体的变构增强剂）增强，两种药物的增强程度取决于细胞。这可能是由于 AMPA 受体的细胞依赖性异质性所致。在这些细胞中，使用膜片钳记录技术记录了 AMPA 受体介导的膜电流。PEPA 也增强了电流反应，并显示出内向整流的 I-V 关系，至少在我们测试的细胞中是这样。我们检查了神经胶质 AMPA 受体激活所刺激的基因表达。为此，对神经胶质细胞培养物中的 mRNA 进行定量 RT-PCR。我们无法检测到我们的培养物中 GDNF mRNA 水平表达的显着增加。我们对其他基因进行了进一步的研究，并制备了其他脑区的胶质细胞，以获得关于胶质细胞AMPA受体的进一步信息。

项目成果

Takezawa,T et al.: "Concept for organ engineering : a reconstruction method of rat ......"Tissue Engineering. 6. 641-650 (2000)

Takezawa,T 等人：“器官工程的概念：大鼠的重建方法……”组织工程。

Sekiguchi M.: "A Novel Allosteric Potentiator of AMPA Receptors"J. Neurosci.. 17. 5760-5771 (1997)

Sekiguchi M.：“AMPA 受体的新型变构增效剂”J。

Nakagawa,T et al.: "Potentiating effects of 4-[2-(phenylsulfonylamino)ethylthio]-2,6-difluoro-……"Neurosci.Res. 35. 217-223 (1999)

Nakakawa, T 等人：“4-[2-(苯磺酰氨基)乙硫基]-2,6-二氟-...的增强作用” Neurosci.Res. 35. 217-223 (1999)

Harada T.: "Light-induced retinal degeneration suppresses developmental progression of flip-to-flop alternative splicing in GluR1"J. Neurosci.. 18. 3336-3343 (1998)

Harada T.：“光诱导的视网膜变性抑制 GluR1 中翻转选择性剪接的发育进程”J。

Takezawa,T et al.: "Concept for organ engineering : a reconstruction method of rat......"Tissue Engineering. 6. 641-650 (2000)

Takezawa,T 等人：“器官工程概念：大鼠的重建方法……”组织工程。