测量分离的线粒体和透化细胞中的线粒体 Ca 2+流出

Statistical analysis of instrumental reproducibility as internal quality control in high-resolution respirometry

Keywords：mitochondria, Ca2+ efflux, NCLX, mPTP, liver 

关键词：线粒体、Ca 2+流出、NCLX、mPTP、肝脏 
作者：Serna Julian DC , Ramos Vitor M , Cabral-Costa Joao V , Vilas-Boas Eloisa A , Amaral Andressa G , Ohya Georgia , da Silva Camille CC , Kowaltowski Alicia J
出版期刊：《Bioenerg Commun》（2022/7/28）

Abstract：

Mitochondrial Ca2+ efflux is essential for mitochondrial and cell Ca2+ homeostasis. Mitochondrial inner membrane Ca2+/H+ and Na+/Li+/Ca2+ (NCLX) exchangers are known today to be plastic transporters, with important roles in physiological responses and pathological states. Until now, however, no consensus protocols were available to measure mitochondrial Ca2+ efflux, and we find that some published protocols may induce mitochondrial permeability transition, underestimating the effects of these exchangers. In this work we describe a method to measure Na+-sensitive and insensitive mitochondrial Ca2+ efflux activity in isolated mitochondria and permeabilized cells using the Ca2+ Green indicator and a fluorimeter. A checklist is provided to avoid artefacts as well as pinpoint adaptations necessary in specific experimental models.

文章摘要：

线粒体 Ca 2+流出对于线粒体和细胞 Ca 2+稳态至关重要。目前已知线粒体内膜 Ca 2+ /H +和 Na + /Li + /Ca 2+ (NCLX) 交换剂是塑料转运体，在生理反应和病理状态中具有重要作用。然而，到目前为止，还没有一致的协议可用于测量线粒体 Ca 2+流出，我们发现一些已发表的协议可能会导致线粒体通透性转变，低估了这些交换剂的影响。在这项工作中，我们描述了一种测量 Na +敏感和不敏感线粒体 Ca的方法使用 Ca 2+ Green 指示剂和荧光计在分离的线粒体和透化细胞中的2+流出活性。提供了一个清单以避免伪影以及特定实验模型中必要的精确调整。

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