Neuroprotection of rat retinal ganglion cells mediated through alpha7 nicotinic acetylcholine receptors.
Neuroprotection of rat retinal ganglion cells mediated through alpha7 nicotinic acetylcholine receptors.
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DOI：
10.1016/j.neuroscience.2013.02.003
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发表时间：
2013-05-01
期刊：
影响因子：
3.3
通讯作者：
Linn CL
中科院分区：
文献类型：
--
作者：
Iwamoto K;Mata D;Linn DM;Linn CL
谷氨酸诱导的兴奋性毒性被认为在中枢神经系统的几种神经退行性疾病中起重要作用。在这项研究中，在培养的成年大鼠视网膜神经元中，使用乙酰胆碱、尼古丁和α 7特异性烟碱乙酰胆碱受体激动剂PNU-282987分析了对谷氨酸诱导的兴奋性毒性的神经保护作用。分离成年Long Evans大鼠视网膜，并使用两步淘选技术从所有其他视网膜组织中分离RGC。一旦分离，RGC在各种药理学条件下培养以证明兴奋性毒性和针对兴奋性毒性的神经保护。三天后，用针对糖蛋白Thy 1.1的抗体对RGC进行免疫染色，计数，并相对于对照未处理条件评估细胞存活。500 µM谷氨酸在成年大鼠分离培养物中的大小RGCs中诱导兴奋性毒性。在与谷氨酸一起培养三天后，大RGC的细胞存活率平均下降了48.16%，而小RGC的细胞存活率平均下降了42.03%。使用特定的谷氨酸受体激动剂和拮抗剂，我们提供的证据表明，兴奋性毒性反应是通过AMPA/KA和NMDA谷氨酸受体介导的凋亡机制。然而，如果在谷氨酸损伤前将细胞与10 µM ACh、100 µM尼古丁或100 nM α7 nAChR激动剂PNU-282987培养1小时，则谷氨酸对所有RGC的兴奋毒性作用均被消除。使用10 nM MLA或α-Bgt的抑制研究支持以下假设，即大鼠RGC对谷氨酸诱导兴奋性毒性的神经保护作用是通过α7 nAChR介导的。在免疫细胞化学研究中，使用抗Thy 1.1和α 7 nAChR亚基的抗体进行的双标记实验表明，大型和小型RGC均含有α 7 nAChR亚基。本研究提供的数据支持ACh和nAChR激动剂通过激活α7 nAChR亚单位对成年大鼠RGC中谷氨酸诱导的兴奋性毒性提供神经保护的假设。这些研究为使用兴奋性毒性体内模型分析特定α7 nAChR激动剂的作用奠定了基础。了解参与大鼠视网膜神经保护的ACh受体类型可能最终导致任何涉及兴奋性毒性的CNS疾病的治疗。
Glutamate-induced excitotoxicity is thought to play an important role in several neurodegenerative diseases in the CNS. In this study, neuroprotection against glutamate-induced excitotoxicity was analyzed using acetylcholine, nicotine and the alpha7 specific nicotinic acetylcholine receptor agonist, PNU-282987, in cultured adult rat retinal neurons. Adult Long Evans rat retinas were dissociated and RGCs were isolated from all other retinal tissue using a two-step panning technique. Once isolated, RGCs were cultured under various pharmacological conditions to demonstrate excitotoxicity and neuroprotection against excitotoxicity. After three days, RGCs were immunostained with antibodies against the glycoprotein, Thy 1.1, counted and cell survival was assessed relative to control untreated conditions. 500 µM glutamate induced excitotoxicity in large and small RGCs in an adult rat dissociated culture. After three days in culture with glutamate, the cell survival of large RGCs decreased by an average of 48.16% while the cell survival of small RGCs decreased by an average of 42.03%. Using specific glutamate receptor agonists and antagonists, we provide evidence that the excitotoxic response was mediated through AMPA/KA and NMDA glutamate receptors through an apoptotic mechanism. However, the excitotoxic effect of glutamate on all RGCs was eliminated if cells were cultured for an hour with 10 µM ACh, 100 µM nicotine or 100 nM of the α7 nAChR agonist, PNU-282987, before the glutamate insult. Inhibition studies using 10 nM MLA or α-Bgt supported the hypothesis that neuroprotection against glutamate-induced excitotoxicity on rat RGCs was mediated through α7 nAChRs. In immunocytochemical studies, double-labeled experiments using antibodies against Thy 1.1 and alpha7 nAChR subunits demonstrated that both large and small RGCs contained alpha7 nAChR subunits. The data presented in this study supports the hypothesis that ACh and nAChR agonists provide neuroprotection against glutamate-induced excitotoxicity in adult rat RGCs through activation of α7 nAChR subunits. These studies lay the groundwork required for analyzing the effect of specific α7 nAChR agonists using in vivo models of excitotoxicity. Understanding the type of ACh receptors involved in neuroprotection in the rat retina could ultimately lead to therapeutic treatment for any CNS disease that involves excitotoxicity.
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影响因子：
4.7
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