急性分离大鼠皮层神经元的方法与体会(1)
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[摘要] 目的：建立急性分离大鼠皮层神经元的方法。方法：切取活脑组织切片，经Protease酶解消化、孵育后，采用酶加机械分离法制备10~16 d鼠龄的大鼠皮层神经元。作形态学观察，用全细胞膜片钳技术记录其电生理学特性。结果：分离出的神经元倒置相差显微镜下形态保持良好，有较长的轴突；用膜片钳技术证实，80%以上细胞可以记录出电活动，其保存了主要的离子通道活性。结论：成功建立了一种经济、简便的适用于膜片钳技术的大鼠皮层神经元急性分离方法。
[关键词] 皮层神经元；急性分离；膜片钳；离子通道
[中图分类号] R965 [文献标识码] A [文章编号] 1673-7210(2011)06(c)-041-03
Study on the separating the cortical neurons of rats by acute enzyme separation method
WANG Zhuang1, LIU Fu2, JI Hui3, TIAN Hua4*
1.Office of Student Work of Adult and Further Education College of Qiqihar Medical University, Qiqihar 161006, China; 2.Office of Basic Medical College of Qiqihar Medical University, Qiqihar 161006, China;3.Department of Anatomy of Qiqihar Medical University, Qiqihar 161006, China;4.Department of Pharmacology of Qiqihar Medical University, Qiqihar 161006, China
[Abstract] Objective: To establish a method which is described for separating the rat cortical neurons. Methods: Cuting a slice of live brain sections, after Protease enzymatic hydrolysis digestion, incubation, the cortical neurons were obtained by a combination of mechanical and enzymatic means from 10-16 day-old rats. Observing by morphology, their electrophysiological properties were recorded by whole-cell patch-clamp recording technique. Results: the difference of the inverted cortical neurons which were obtained by separation kept good under microscope. They each had a long axon. Proved by the patch-clamp recording technique, more than 80% cells could be recorded their electric activity, which preserved the ion channel activity. Conclusion: An economic and simple method was established, which is described for separating the rat cortical neurons by the patch-clamp recording technique.
[Key words] Cortical neurons; Acute isolation; Patch clamp; Ion channel
近年来 ......
[摘要] 目的：建立急性分离大鼠皮层神经元的方法。方法：切取活脑组织切片，经Protease酶解消化、孵育后，采用酶加机械分离法制备10~16 d鼠龄的大鼠皮层神经元。作形态学观察，用全细胞膜片钳技术记录其电生理学特性。结果：分离出的神经元倒置相差显微镜下形态保持良好，有较长的轴突；用膜片钳技术证实，80%以上细胞可以记录出电活动，其保存了主要的离子通道活性。结论：成功建立了一种经济、简便的适用于膜片钳技术的大鼠皮层神经元急性分离方法。
[关键词] 皮层神经元；急性分离；膜片钳；离子通道
[中图分类号] R965 [文献标识码] A [文章编号] 1673-7210(2011)06(c)-041-03
Study on the separating the cortical neurons of rats by acute enzyme separation method
WANG Zhuang1, LIU Fu2, JI Hui3, TIAN Hua4*
1.Office of Student Work of Adult and Further Education College of Qiqihar Medical University, Qiqihar 161006, China; 2.Office of Basic Medical College of Qiqihar Medical University, Qiqihar 161006, China;3.Department of Anatomy of Qiqihar Medical University, Qiqihar 161006, China;4.Department of Pharmacology of Qiqihar Medical University, Qiqihar 161006, China
[Abstract] Objective: To establish a method which is described for separating the rat cortical neurons. Methods: Cuting a slice of live brain sections, after Protease enzymatic hydrolysis digestion, incubation, the cortical neurons were obtained by a combination of mechanical and enzymatic means from 10-16 day-old rats. Observing by morphology, their electrophysiological properties were recorded by whole-cell patch-clamp recording technique. Results: the difference of the inverted cortical neurons which were obtained by separation kept good under microscope. They each had a long axon. Proved by the patch-clamp recording technique, more than 80% cells could be recorded their electric activity, which preserved the ion channel activity. Conclusion: An economic and simple method was established, which is described for separating the rat cortical neurons by the patch-clamp recording technique.
[Key words] Cortical neurons; Acute isolation; Patch clamp; Ion channel
近年来 ......
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