可视法脑片膜片钳实验技术的建立及大鼠前庭内侧核神经元毒蕈碱样胆碱能受体电生理特性的研究

发布时间：2018-11-07 17:18

【摘要】： 【目的】建立大鼠前庭内侧核脑片可视法膜片钳实验技术,探讨在前庭内侧核神经元(medial vestibular nucleus neurons,MVNn)中毒蕈碱样胆碱能受体(M受体)介导电流的特性。【方法】应用红外微分干涉相差(infrared differential interference contrast,IR-DIC)显微镜结合电荷耦合式摄像机(charge coupled device-camera,CCD-camera)系统,可视法膜片钳全细胞记录模式下检测前庭内侧核神经元毒蕈碱样胆碱能受体介导电流的特性。【结果】可视法脑片膜片钳技术可对神经元直接进行准确定位以及健康细胞的筛选。在MVNn中,毒蕈碱引发的M受体去极化效应是由钾电导的减少所介导,并证明所抑制的离子电流是漏钾电流。【结论】可视法脑片膜片钳实验技术克服了盲法脑片膜片钳技术的缺陷,大大提高了对神经元封接的成功率。通过对MNVn中M受体通道电流性质的分析,从而进一步揭示毒蕈碱样胆碱能机制在MVNn的兴奋性调节作用,并为临床抗胆碱药物的应用提供新的思路。
[Abstract]:[objective] to establish a visual patch clamp technique in the medial vestibular nucleus of rats to investigate the characteristics of muscarinic cholinergic receptor (M receptor) mediated currents in the neurons of the medial vestibular nucleus (medial vestibular nucleus neurons,MVNn). [methods] an infrared differential interference phase difference (infrared differential interference contrast,IR-DIC) microscope combined with a charge-coupled camera (charge coupled device-camera,CCD-camera) system was used. The characteristics of muscarinic cholinergic receptor-mediated currents in medial vestibular nucleus neurons were detected by visual patch clamp whole-cell recording mode. [results] the visual patch clamp technique can directly locate neurons and screen healthy cells. In MVNn, the depolarization of M receptor induced by muscarinic acid is mediated by the reduction of potassium conductance, and it is proved that the inhibited ion current is potassium leakage current. [conclusion] the visual patch-clamp technique overcomes the defect of blind patch-clamp technique and improves the success rate of neuron sealing. By analyzing the current properties of M-receptor channel in MNVn, the excitatory regulation of muscarinic cholinergic mechanism in MVNn was further revealed, and a new idea for clinical application of anticholine drugs was provided.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R33

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