Eugene and Marilyn Glick Eye Institute, Department of Ophthalmology, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
STAR Protoc. 2021 May 12;2(2):100533. doi: 10.1016/j.xpro.2021.100533. eCollection 2021 Jun 18.
Mitochondrial metabolism is a critical mechanism that is deregulated in numerous retinal diseases. Here, we elaborate a protocol to quantify oxygen consumption rate as a measure of mitochondrial respiration directly from mouse retinal tissue pieces. Our procedure combines the use of Seahorse extracellular flux technology and retinal tissue isolation and is robustly reproducible under different treatment conditions. This protocol allows direct assessment of mitochondrial function in response to drug treatments or genetic manipulation in mouse models. For complete details on the use and execution of this protocol, please refer to Shetty et al. (2020), Sardar Pasha et al. (2021), Kooragayala et al. (2015), and Joyal et al. (2016).
线粒体代谢是许多视网膜疾病中失调的关键机制。在这里，我们详细介绍了一种从鼠视网膜组织块中直接定量氧消耗率作为线粒体呼吸测量的方案。我们的程序结合了 Seahorse 细胞外通量技术和视网膜组织分离的使用，在不同的处理条件下具有很强的重现性。该方案允许直接评估药物处理或遗传操作后对小鼠模型中线粒体功能的影响。有关该方案使用和执行的完整详细信息，请参阅 Shetty 等人（2020 年）、Sardar Pasha 等人（2021 年）、Kooragayala 等人（2015 年）和 Joyal 等人（2016 年）。
