一种改进的适用于膜片钳记录的成年大鼠海马神经元急性分离法(英文)  

An improved method for acute isolation of neurons from the hippocampus of adult rats suitable for patch-clamping study

作　　者：李晓明[1] 李建国[2] 杨建明[2] 胡平[2] 李晓文[2] 王颖[2] 覃鲁宁[2] 高天明[1] 

机构地区：[1]第一军医大学解剖学教研室 [2]第一军医大学生理学教研室

出　　处：《生理学报》2004年第1期112-117,共6页Acta Physiologica Sinica

基　　金：This work is supposed by grants from Outstanding Young Scientists Program of NSFC(30125013);Outstanding Young Scientists Program of PLA(01J009);NSFC/RGC Joint Research Scheme(30218004);Natural Science Foundation of Guangdong Province (010639);Cheung

摘　　要：本文建立了一种快速、可靠的急性分离成年大鼠海马神经细胞的方法。此法可将实验大鼠的年龄提高到500 d以上,体重300 g以上;不损伤神经细胞膜的电学特性;形态上有差异的细胞易于分辨。用膜片钳技术的单通道和全细胞模式证实,在本实验条件下,约95％左右的健康细胞均能形成高阻抗封接,并成功地记录了电压依赖性钾、钠、钙通道,外向整流氯通道和大电导的钙激活钾通道电流。An improved method is described for fast and reliable isolation of neurons from hippocampus of adult rats by a combination of mechanical and enzymatic means. The procedure allows the isolation of neurons from 500-600-d-old rats (over 300 g), preserving the proximal dendritic structure without impairing the electrical characteristics of the cells. Morphologically distinct neurons can be recognized. Using cell-attached, inside-out and whole-cell configurations of patch clamp technique, it was shown that the enzymatically isolated neurons in hippocampus from rats weighing more than 300 g exhibited voltage-gated calcium, sodium and potassium currents, outwardly rectifying chloride channel and large conductance Ca2+-activated potassium channel currents. Approximately, 95% of healthy cells allowed the formation of giga-ohm seals.

关 键 词：神经元 膜片钳 海马 大鼠 

分 类 号：R33[医药卫生—人体生理学]