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目的:利用成年5-HT3AR-BACEGFP转基因小鼠,研究5-羟色胺3A受体(5-HT3AR)在海马中间神经元中的分布情况。方法:成年5-HT3AR-BACEGFP转基因小鼠经心脏灌注固定后,利用免疫荧光双标记方法,并结合激光共焦显微镜扫描技术,观察5-HT3AR在成年5-HT3AR-BACEGFP转基因小鼠海马中不同中间神经元内的表达和分布情况。结果:5-HT3AR在成年小鼠整个海马中都有分布,且主要在CA1区、CA2/CA3区和齿状回有大量5-HT3AR免疫阳性细胞;激光共聚焦显微镜下观察到5-HT3AR阳性产物在细胞核、细胞浆和树突上均有表达;免疫荧光双标实验结果表明5-HT3AR阳性产物在CB(calbindin),CR(calretinin),Reelin,Som(somatostatin),NPY(neuropeptide Y)和VIP(vasoactive intestinal peptide)免疫阳性神经元中表达,但在PV(parvalbumin)免疫阳性神经元中不表达。定量结果显示:几乎所有的VIP阳性神经元均表达5-HT3AR阳性,约3/4的CR阳性神经元表达5-HT3AR,约1/2的CB、Reelin、NPY和Som阳性神经元表达5-HT3AR阳性;约1/4的5-HT3AR阳性神经元中表达Reelin,1/5的表达Som,5-HT3AR/CB和5-HT3AR/CR双标神经元各占5-HT3AR阳性神经元的1/10左右。结论:5-HT3AR-BACEGFP转基因小鼠能够作为研究海马中5-HT3AR功能及其在中间神经元中的作用机制研究的工具鼠。
OBJECTIVE: To study the distribution of 5-hydroxytryptamine 3A receptor (5-HT3AR) in the hippocampal interneurons using adult 5-HT3AR-BACEGFP transgenic mice. METHODS: Adult 5-HT3AR-BACEGFP transgenic mice were perfused with cardioplegia. The immunofluorescence double labeling and laser scanning confocal microscopy were used to observe the difference of 5-HT3AR in the hippocampus of adult 5-HT3AR-BACEGFP transgenic mice Interneurons expression and distribution. Results: 5-HT3AR was distributed in the whole hippocampus of adult mice and there was a large number of 5-HT3AR immunoreactive cells mainly in CA1, CA2 / CA3 and dentate. 5-HT3AR positive cells were observed under confocal laser scanning microscope The results of double immunofluorescence staining showed that the positive products of 5-HT3AR in calbindin, calretinin, reelin, somatostatin, NPY (neuropeptide Y) and VIP (vasoactive intestinal peptide) immunoreactive neurons, but not in parvalbumin immunoreactive neurons. Quantitative results showed that almost all of the VIP positive neurons expressed 5-HT3AR, about 3/4 CR positive neurons expressed 5-HT3AR, about 1/2 CB, Reelin, NPY and Som positive neurons expressed 5- HT3AR positive; 5% of 5-HT3AR positive neurons of Reelon, 1/5 Som, 5-HT3AR / CB and 5-HT3AR / CR double-labeled neurons / 10 or so. CONCLUSIONS: 5-HT3AR-BACEGFP transgenic mice can be used as a tool to study the role of 5-HT3AR in the hippocampus and its role in interneurons.