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首页> 外文期刊>日本眼科學會雜誌 >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 fors-kolin. 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 GluR 1 to 4 and microtubuleassociated protein (MAP) 2, which is a neuronal marker. Glutamate-induced Ca~(2+) influx was measured with fura-2 acet-oxymethyl ester fluorescence.
机译:目的:确定培养的视网膜神经节细胞(RGC)中是否表达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:采用改良的两步淘选法从离体大鼠视网膜细胞(出生后第6-8天)中纯化RGC,并在含有神经营养因子和Fors-kolin的无血清培养基中培养。使用抗AMPAR亚基GluR 1至4和微管相关蛋白(MAP)2的特异性抗体在体外第1、3和7天(1 DIV,3 DIV和7 DIV)对培养的RGC进行免疫组织化学,这是一种神经元标记。谷氨酸诱导的Ca〜(2+)流入量用fura-2乙酰氧基甲基酯荧光法测定。

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