Bergmann Corinna, Basta Dietmar, Ptok Martin, Ernst Arne
Department of Biology and Biochemistry, University of Potsdam, Germany.
Eur Arch Otorhinolaryngol. 2005 Dec;262(12):965-8. doi: 10.1007/s00405-005-0932-3. Epub 2005 Jun 18.
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.
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。