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成熟下丘神经元链霉蛋白酶解离的质量

Quality of pronase dissociation of mature inferior colliculus neurons.

作者信息

Bergmann Corinna, Basta Dietmar, Ptok Martin, Ernst Arne

机构信息

Department of Biology and Biochemistry, University of Potsdam, Germany.

出版信息

Eur Arch Otorhinolaryngol. 2005 Dec;262(12):965-8. doi: 10.1007/s00405-005-0932-3. Epub 2005 Jun 18.

Abstract

One major advantage of acutely dissociated inferior colliculus (IC) neurons in electrophysiological investigations is their complete isolation from the surrounding cellular network. In this way, patch-clamp recordings can be performed under controlled conditions to study membrane properties of IC neurons in more detail. The aim of the present study was to adapt a dissociation method for immature IC neurons to the highly sensitive, fragile and vulnerable mature IC neurons of mammals (mice). The modification of a pronase-based dissociation protocol with respect to concentration, incubation time and handling (trituration) of the cells yielded intact, live IC neurons with a clean cell surface so that they were well suited for further electrophysiological investigations in our study. The largely modified dissociation protocol is described in detail and critically discussed.

摘要

在电生理研究中,急性分离的下丘(IC)神经元的一个主要优点是它们与周围细胞网络完全隔离。通过这种方式,可以在受控条件下进行膜片钳记录,以更详细地研究IC神经元的膜特性。本研究的目的是将一种适用于未成熟IC神经元的解离方法应用于哺乳动物(小鼠)高度敏感、脆弱且易受损的成熟IC神经元。对基于链霉蛋白酶的解离方案在细胞浓度、孵育时间和处理(研磨)方面进行修改后,得到了具有干净细胞表面的完整活IC神经元,使其非常适合我们研究中的进一步电生理研究。本文详细描述并批判性地讨论了经过大量修改的解离方案。

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