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目的:检测大鼠前庭终器中毒蕈碱样乙酰胆碱受体(mAChRs)m1~m5亚基的表达。方法:根据GeneBank公布的大鼠mAChRm1~m5亚基的mRNA序列设计特异性引物,再运用RT-PCR方法检测大鼠前庭终器中编码mAChRm1~m5亚基的mRNA的表达,之后行PCR产物直接测序。以大鼠脑干组织提取RNA为阳性对照,以解剖液为阴性对照。结果:扩增出标志mAChRm1~m5亚基表达的PCR产物,并经测序证实与预计扩增的cDNA片段序列一致。结论:在大鼠前庭终器组织中有mAChRm1~m5亚基的表达,为乙酰胆碱作为大鼠外周前庭系统神经递质提供了最直接的证据。
Objective: To detect the expression of m1 ~ m5 subunit of muscarinic acetylcholine receptor (mAChRs) in the vestibular end-organ in rats. Methods: The specific primers were designed according to the mRNA sequences of mAChRm1 ~ m5 subunits published by GeneBank. The mRNA expression of mAChRm1 ~ m5 subunits in the vestibular terminal of rat was detected by RT-PCR. The PCR products were directly Sequencing. RNA was extracted from rat brainstem tissue as positive control and anatomical solution as negative control. Results: The PCR products of mAChRm1 ~ m5 subunits were amplified and confirmed by sequencing. The PCR products were consistent with the predicted cDNA fragments. CONCLUSION: The expression of mAChRm1 ~ m5 subunit in the end-stage vestibular tissue of rat provides the most direct evidence for acetylcholine as the neurotransmitter in rat peripheral vestibular system.