Measurement of mitochondrial respiration in permeabilized fish gills
Measurement of mitochondrial respiration in permeabilized fish gills
复制标题
DOI：
10.1242/jeb.216762
复制
发表时间：
2020-02-01
影响因子：
2.8
通讯作者：
Metcalfe, Neil B.
中科院分区：
文献类型：
--
作者：
Dawson, Neal J.;Millet, Caroline;Metcalfe, Neil B.
鱼鳃的生理研究传统上集中在鳃的两个主要功能：气体交换和离子调节。富含线粒体的细胞（MRCs）主要存在于鱼类的鳃丝中，并且被认为增殖以增加鳃响应环境诱导的渗透挑战的离子调节能力。然而，令人惊讶的是，很少有人注意到鱼鳃内线粒体的代谢功能。在这里，我们描述并验证了一个简单的协议，鱼鳃渗透和随后的线粒体呼吸速率在体外测量。我们的方案只需要小的组织样本（8毫克），利用鱼鳃的天然结构，不需要鳃组织的机械分离（因此相对快速执行），并产生准确和高度可重复的呼吸率测量。这为研究各种鱼类的鳃线粒体功能提供了巨大的潜力。
Physiological investigations of fish gills have traditionally centred on the two principal functions of the gills: gas exchange and ion regulation. Mitochondrion-rich cells (MRCs) are primarily found within the gill filaments of fish, and are thought to proliferate in order to increase the ionoregulatory capacity of the gill in response to environmentally induced osmotic challenges. However, surprisingly little attention has been paid to the metabolic function of mitochondria within fish gills. Here, we describe and validate a simple protocol for the permeabilization of fish gills and subsequent measurement of mitochondrial respiration rates in vitro. Our protocol requires only small tissue samples (8 mg), exploits the natural structure of fish gills, does not require mechanical separation of the gill tissue (so is relatively quick to perform), and yields accurate and highly reproducible measurements of respiration rates. It offers great potential for the study of mitochondrial function in gills over a wide range of fish sizes and species.