星形胶质细胞毒蕈碱样乙酰胆碱受体M1、M3、M5亚型基因克隆及序列分析

Sequence analysis of muscarinic receptors M1, M3 and M5 expression by cultured rat astrocyte cells

目的克隆胶质细胞M1、M3、M5受体亚型基因序列,比较胶质细胞和神经细胞M1、M3、M5受体亚型基因序列、蛋白质序列的差异。方法根据神经细胞M1、M3、M5受体基因序列全长设计特异性探针,采用RT-PCR方法扩增胶质细胞M1、M3、M5受体亚型基因序列,并对其进行克隆测序。结果测序得到胶质细胞M1、M3、M5受体亚型基因序列,与神经细胞比较,M1受体差异碱基4个,发生氨基酸改变的有1个;M3受体差异碱基有8个,发生氨基酸改变的位点有4个;M5受体差异碱基1个,发生氨基酸改变的有1个。结论胶质细胞与神经细胞M1、M3、M5受体亚型在基因和蛋白序列上具有明显差异。

Aim To clone the muscarinic receptors M1, M3 and M5 sequences of astrocyte cells,and compare the gene and protein sequences with those of neurons. Methods Specific primers were designed to clone the M1, M3 and M5 sequences of astrocyte cells by RT-PCR according to those of neurons, then sequenced the sequences. Result By comparing the M1, M3 and M5 sequences expression by astrocyte cells with those by neurons and we found four, eight,and one different bases and one, four and one different amino acids in M 1, M3 and M5 between astrocyte cells and neurons respectively. Conclusions The gene and protein sequence differences are evident in M1, M3 and M5 between astrocyte cells and neurons.