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首页> 外文期刊>Japanese Journal of Ophthalmology >Expression of AMPA receptor subunit proteins in purified retinal ganglion cells.
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Expression of AMPA receptor subunit proteins in purified retinal ganglion cells.

机译：AMPA受体亚基蛋白在纯化的视网膜神经节细胞中的表达。

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摘要

PURPOSE: To determine whether alpha-amino-3-hydroxy-5-methylisoxazole-4-propioate (AMPA) receptor (AMPAR) subunit proteins are expressed in cultured retinal ganglion cells (RGCs). METHODS: RGCs were purified from dissociated rat retinal cells (postnatal days 6-8), using a modified two-step panning method and cultured in serum-free medium containing neurotrophic factors and forskolin. Immunohistochemistry was performed on cultured RGCs on days 1, 3, and 7 in vitro (1 DIV, 3 DIV, and 7 DIV) using specific antibodies against AMPAR subunits GluR1 to 4 and microtubule-associated protein (MAP) 2, which is a neuronal marker. Glutamate-induced Ca(2+) influx was measured with fura-2 acetoxymethyl ester fluorescence. RESULTS: GluR1 to 4 proteins were expressed in the cell body of RGCs on 1 DIV. RGCs showed strong GluR1 to 4 immunoreactivity in both cell bodies and processes on 3 DIV and 7 DIV, with the gradual spreading of expression and the growth of processes. At all time points examined, GluR2 immunoreactivity was equal to that of the other subunits. Accumulation of intracellular Ca(2+) levels in RGCs induced by glutamate occurred equally on both 3 DIV and 7 DIV. CONCLUSION: All AMPAR subunits are almost equally expressed in cultured RGCs.
机译：目的：确定是否在培养的视网膜神经节细胞（RGCs）中表达α-氨基-3-羟基-5-羟基-5-甲基异恶唑-4-丙酸酯（AMPA）受体（AMPAR）亚基蛋白。方法：使用改良的两步淘选法从分离的大鼠视网膜细胞（出生后6-8天）中纯化RGC，并在含有神经营养因子和毛喉素的无血清培养基中培养。使用抗AMPAR亚基GluR1至4和微管相关蛋白（MAP）2的特异性抗体在体外培养的RGC上第1、3和7天（1 DIV，3 DIV和7 DIV）进行免疫组织化学标记。谷氨酸诱导的Ca（2+）涌入用fura-2乙酰氧基甲基酯荧光测量。结果：在1 DIV上，RGC的细胞体中表达了GluR1至4蛋白。 RGC在3 DIV和7 DIV的细胞体和过程中均显示出强大的GluR1至4免疫反应性，并且表达逐渐扩散且过程不断增长。在检查的所有时间点，GluR2免疫反应性均与其他亚基相同。谷氨酸诱导的RGC中细胞内Ca（2+）的积累在3 DIV和7 DIV上均等发生。结论：所有AMPAR亚基在培养的RGC中几乎均等表达。

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