急性分离Meynert核团神经元方法及其应用

The method for dissociating acutely neurons of Meynert nucleus and its application

目的建立急性分离Meynert核团神经元方法。方法切取活脑组织切片,经Protease酶解消化、孵育后,机械分离Meynert核团神经元。作形态学观察,通过单细胞膜片钳技术记录电活动。结果分离出的Meynert核团神经元,倒置相差显微镜下形态保持良好,突起较完整。90%以上细胞可以记录出电活动。结论本方法可以获取活的Meynert核团单个神经元,用于单细胞膜片钳、激光共聚焦、流式细胞术等研究。

Objective To establish the method for acutely dissociating neurons of Meynert nucleus in rat to provide the technique for basic research of senile diseases. Methods Brain slices obtained were digested by protease enzyme, then incubated in extracellular fluid. Meynert nucleus was obtained and dissociated to observe their morphology and record electric activity by patch clamp technique. Results Under inverted microscope, cellular membrane of neurons of nucleus basalis of Meynert （nbM） was smooth and glossy, its soma were fusiform, or cone, or ellipse in shape. Electric activity was showed in more than 90% neurons. Conclusions This method was better technique for obtaining neurons of Meynert nucleus, which can be used in many experiments, such as the unicell patch clamp, laser scanning confocal microscope, flow cytometer and so on.