恒河猴视网膜中的烟碱型乙酰胆碱受体亚基
Nicotinic acetylcholine receptor subunits in rhesus monkey retina.
作者信息
Liu Ji, McGlinn Alice M, Fernandes Alcides, Milam Ann H, Strang Christianne E, Andison Margot E, Lindstrom Jon M, Keyser Kent T, Stone Richard A
机构信息
Department of Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, Pennsylvania 19104-6075, USA.
出版信息
Invest Ophthalmol Vis Sci. 2009 Mar;50(3):1408-15. doi: 10.1167/iovs.08-2398. Epub 2008 Oct 24.
PURPOSE
The purpose of this study was to detect and establish the cellular localizations of nicotinic acetylcholine receptor (nAChR) subunits in Rhesus monkey retina.
METHODS
Retinas were dissected from the eyes of monkeys killed after unrelated experiments. RNA was extracted and analyzed by RT-PCR, using primers designed against human sequences of alpha3-alpha7, alpha9, and beta2-beta4 nAChR subunits. The RT-PCR products were separated by gel electrophoresis and sequenced. Frozen sections of postmortem fixed monkey eyes were immunolabeled with well-characterized and specific monoclonal antibodies against the alpha3, alpha4, alpha6, alpha7, beta2, or beta4 nAChR subunits and visualized with fluorescence labeling.
RESULTS
Products of the predicted size for the alpha3-alpha7, alpha9, and beta2-beta4 nAChR subunits were detected by RT-PCR in Rhesus monkey retina. Homology between transcripts from monkey retina and human nucleotide sequences ranged from 93 to 99%. Immunohistochemical studies demonstrated that neurons in various cell layers of monkey retina expressed alpha3, alpha4, alpha7, or beta2 nAChR subunits and cells with the morphology of microglia were immunoreactive for the alpha6 or beta4 nAChR subunits.
CONCLUSIONS
nAChR subunits are expressed in the monkey retina and localize to diverse retinal neurons as well as putative microglia. Besides mediating visual processing, retinal nAChRs may influence refractive development and ocular pathologies such as neovascularization.
目的
本研究旨在检测并确定烟碱型乙酰胆碱受体（nAChR）亚基在恒河猴视网膜中的细胞定位。
方法
从 unrelated experiments 后处死的猴子眼中取出视网膜。提取 RNA 并通过 RT-PCR 进行分析，使用针对人α3-α7、α9以及β2-β4 nAChR 亚基序列设计的引物。RT-PCR 产物经凝胶电泳分离并测序。对死后固定的猴眼冰冻切片用针对α3、α4、α6、α7、β2或β4 nAChR 亚基的特性明确且特异的单克隆抗体进行免疫标记，并用荧光标记进行可视化观察。
结果
通过 RT-PCR 在恒河猴视网膜中检测到了α3-α7、α9以及β2-β4 nAChR 亚基预测大小的产物。猴视网膜转录本与人核苷酸序列之间的同源性在 93%至 99%之间。免疫组织化学研究表明，猴视网膜各细胞层中的神经元表达α3、α4、α7或β2 nAChR 亚基，具有小胶质细胞形态的细胞对α6或β4 nAChR 亚基呈免疫反应性。
结论
nAChR 亚基在猴视网膜中表达，并定位于多种视网膜神经元以及假定的小胶质细胞。除了介导视觉处理外，视网膜 nAChRs 可能影响屈光发育和眼部病变，如新生血管形成。
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