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Neurophysiological study for excitatory amino acids-induced neurotoxicity and apoptosis in cultured retinal neurons.

培养的视网膜神经元中兴奋性氨基酸诱导的神经毒性和细胞凋亡的神经生理学研究。

基本信息

批准号：
09671801
负责人：
MISHIMA Hiromu
金额：
$ 2.05万
依托单位：
HIROSHIMA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

1. Protection of vasoactive intestinal peptide (VIP) against glutamate-induced neurotoxicity in cultured retinal neurons.VIP (10 nM - 1 muM) was attenuated glutamate-induced neurotoxicity in a concentration-dependent manner. This protective effect was antagonized by VIP6-28, a VIP recptor antagonist, and H-89, a protein kinase A (PKA) inhibitor. VIP did not affect glutamate-induced current and Ca^<2+> influx. VIP is suggested to act as a protective factor against glutamate-induced neurotoxicity of retinal neurons by elevating the cAMP level via VIP receptors. Activated PKA might have an effect downstream of the Ca^<2+> influx in glutamate neurotoxicity.2. Protection of pituitary adenylate cyclase activating peptide (PACAP) against glutamate-induced neurotoxicity in cultured retinal neurons.PACAP27 (10 nM - 1 muM) and PACAP38 (10 nM - 1 muM) were attenuated glutamate-induced neurotoxicity in a concentration-dependent manner. Activation of mitogen activated protein (MAP) kinase by PACAPs was inhibited by H-89. PACAPs are suggested to act as neuroprotective factors against glutamate-induced neurotoxicity by activating MAP kinase, which is due to PKA activation via PACAP receptors.3. Rat retinal ganglion cells (RGCs) culture enriched with the magnetic cell sorter (MACS).We demonstrated a new RGC-culture technique using MACS.It enriched RGCs from 0.55 % to 31.0 % of all retinal neurons and enhanced the survival of enriched RGCs for over 2 weeks. The whole-cell patch clamp evaluation was suggested that three ionotropic glutamate receptor subtypes such as NMDA, kainate and AMIPA receptors existed on MACS-separated RGCs. This culture system is thought to be an appropriate model for studying glutamate-induced neurotoxicity in RGC.

1.血管活性肠肽（VIP）对谷氨酸诱导的视网膜神经元毒性的保护作用VIP（10 nM - 1 μ M）以浓度依赖性方式减轻谷氨酸诱导的神经毒性。这种保护作用可被VIP受体拮抗剂VIP 6 -28和蛋白激酶A（PKA）抑制剂H-89所拮抗。VIP不影响谷氨酸诱导电流和Ca^2+内流。提示VIP通过VIP受体升高cAMP水平，对谷氨酸诱导的视网膜神经元毒性起保护作用。激活的PKA可能在谷氨酸神经毒性的Ca^<2+>内流下游起作用.垂体腺苷酸环化酶激活肽（PACAP）对谷氨酸诱导的视网膜神经元毒性的保护作用PACAP 27（10 nM - 1 μ M）和PACAP 38（10 nM - 1 μ M）以浓度依赖性方式减弱谷氨酸诱导的神经毒性。H-89可抑制PACAPs对丝裂原活化蛋白（MAP）激酶的激活。PACAP可能通过激活MAP激酶而发挥神经保护作用，而MAP激酶的激活是通过PACAP受体激活PKA实现的.用磁性细胞分选仪（MACS）富集大鼠视网膜神经节细胞（RGCs）培养，我们证明了一种新的RGCs培养技术，MACS富集的RGCs从所有视网膜神经元的0.55%增加到31.0%，并提高富集的RGCs的存活超过2周。全细胞膜片钳技术检测结果表明，MACS分离的RGCs上存在NMDA、红藻氨酸和AMIPA三种离子型谷氨酸受体亚型。该培养系统被认为是研究谷氨酸诱导的RGC神经毒性的合适模型。