培养鼠视网膜神经细胞的腺苷/腺苷酸受体研究

The research of Adenosine/Purinergic receptors on cultured ratsretinal neurons

目的 检测体外培养鼠视网膜神经细胞上腺苷/腺苷酸受体的存在。方法 先进行兔视网膜M(?)ller细胞培养,而后依此细胞作基质培养鼠视网膜神经细胞,应用自由钙指示剂Fura－2AM及AHGUS－100系统,监测不同条件下细胞内自由钙的变化,从而确定腺苷/腺苷酸受体的存在。 结果 1.0μmol/L ATP可引起多量神经细胞内自由钙的变化,A2,P2x,P2y受体激动剂在不同浓度下也各自具有类似的作用,此作用不能被电压依赖性钙通道阻滞剂所抑制。结论培养的视网膜神经细胞上具有A2,P2x,P2y的受体,表明ATP及去磷酸代谢物可能在视网膜功能活动中具有重要的神经介质功能。

Objective 1.To examine the possibility of eulturing tremendous rats retinal neurons in vitro and to identify the cell types. 2. To determine the presence of Adenosine/Purinergie receptors on cultured rats retinal neurons,and how the ATP and its metabolie products may play a very important role in the retinal nervous signal current, glia-neuron interactions and in retinal pathology. Method The M(?)ller cells were cultured from adult rabbit retina at first, then , rat retinal neurons were cultured on rabbit retinal M(?)ller cells. Intracellular calcium concentration was monitored by a videospetro-fluorometry ( ARGUS 100/CA) using Fura-2AM-a calcium indictor, calcium transient following ATP and its analogue administration were observed with or withoutreceptor antagonist. Result ATP at 1.0μmol/L induced cytosolie calcium transient increase in many neurons, A2 P2x,P2yreceptor agonist at different concentration could increase the cytosolie calcium in some neurons respectively. In contrast, A1 agonisthas no such function. The cytosolic calcium increased function was not blocked under the presence of Nifedipine-a voltage dependent calcium channel block, A2-antagonist DMPX could partly block the calcium increase in neurons induced by ATP, but abolish the calcium changes induced by A2 receptor agonist. Conclusion Rats retinal neurons possess A2,P2x,P2y receptors.