烟碱型乙酰胆碱受体在离体海马小胶质细胞上的表达与定位

Expression and localization of α7-nicotinic acetylcholine receptors on rat hippocampal microglial cells

目的探讨烟碱型乙酰胆碱受体（α7-nAChR）在体外培养海马小胶质细胞上的表达和定位。方法取新生SD大鼠的海马组织，进行胶质细胞的混合培养，然后分离纯化小胶质细胞，采用免疫荧光染色检测CD11b／c进行小胶质细胞的鉴定：采用RT—PCR和免疫荧光双标分别检测α7-nAChR在小胶质细胞中的表达和定位。结果免疫荧光染色显示分离纯化的细胞表达小胶质细胞特异性抗体CD11b／c；RT—PCR能扩增出长度为450bp的α7-nAChR目的条带．免疫荧光双标检测显示小胶质细胞CD11b／c和α7-nAChR染色阳性，分别呈红色和绿色荧光，叠加后呈棕黄色，且细胞膜荧光信号较强。结论α7-AChR在体外培养的海马小胶质细胞中能正常表达．定位在细胞膜的功能性α7-nAChR较丰富。

Objective To investigate the expression and distribution of α7-nicotinic acetylcholine receptors （α7-nAChR） on rat hippocampal microglial cells. Mehtods Mixed primary glial cells obtained from the cerebral cortex of 1-day-old rats were cultured for 7-9 days, and the microglial cells were purified. The expression of α7-nAChR at the protein and mRNA levels on the microglia cells was detected using double immunolabeling with anti-CD11b/c antibody and RT-PCR, respectively. Results The resting microglial cells harvested from mixed primary glial culture presented with ramified surface covered with spines, which may serve as a unique feature for identifying microglial cells. A band of 450-bp corresponding to α7-nAChR was specifically amplified by RT-PCR. Double immunolabeling showed colocalization of α7-nAChR and CD11b/c on the cultured hippocampal mcrioglial cells. Conclusion α7-nAChR can be normally expressed in rat hippocampal microglial cells in in vitro culture.