牛视网膜离体灌注-视网膜功能药理研究的灵敏工具

Matthias Lüke , Marco Weiergräber , Carl Brand , Siarhei A. Siapich , Mohammed Banat , Jürgen Hescheler , Christoph Lüke , Toni Schneider
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引用次数: 50

摘要

视网膜电图(ERG)的分离牛视网膜可作为视网膜活动的一个证明标准。它被用作一种敏感的药理学工具,用于测试应用药物和毒素对光感受器的影响,以及有助于产生b波的高阶神经元。分离并脱离下层色素上皮后,将部分视网膜装入封闭腔内,用营养液灌注。经验优化了营养液及其成分的流速、培养温度和光照强度,以获得最大的b波振幅。在这些条件下,一个可重复的,高分辨率的ERG可以稳定地记录超过10小时,并且足够的氧合被认为是持久稳定性的先决条件。在营养液中添加L(+)谷氨酸对b波振幅不再有利。一种著名的氧化磷酸化抑制剂(KCN)和电压门控制的Ca2+通道拮抗剂(isradipine, ω-conotoxin-GVIA和NiCl2)被用来证明测试系统的有效性。从分离和灌注的牛视网膜中记录的ERG可以作为一个有价值的神经网络生理模型,其中与视网膜信号传导和代谢相关的重要问题可以被研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The isolated perfused bovine retina—A sensitive tool for pharmacological research on retinal function

The electroretinogram (ERG) of the isolated bovine retina serves as a proven criterion of retinal activity. It is used as a sensitive pharmacological tool for testing effects of applied drugs and toxins on photoreceptors, and higher order neurons that contribute to the generation of the b-wave. Following isolation and detachment from the underlying pigment epithelium, part of the retina was mounted into a closed chamber and perfused by a nutrient solution. Flow rate of the nutrient solution and its ingredients, incubation temperature and light intensity were optimised empirically to achieve a maximum b-wave amplitude. Under these conditions, a reproducible, high-resolution ERG can be stably recorded for more than 10 h with sufficient oxygenation found to be a prerequisite for the long-lasting stability. Addition of L(+)glutamate to the nutrient solutions was not anymore beneficial for the b-wave amplitude. A well-known inhibitor of oxidative phosphorylation (KCN) and antagonists of voltage-gated Ca2+ channels (isradipine, ω-conotoxin-GVIA and NiCl2) were used to prove the validity of the test system. The recording of the ERG from the isolated and perfused bovine retina serves as a valuable physiological model for a neuronal network in which important questions related to the retinal signalling and metabolism can be investigated.

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