大鼠冰冻结肠组织中线粒体呼吸功能的测量方案。

Protocol for measuring respiratory function of mitochondria in frozen colon tissue from rats.

机构信息

Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 1N4, Canada; Alberta Children's Hospital Research Institute, University of Calgary, Calgary, AB T2N 1N4, Canada.

出版信息

STAR Protoc. 2023 Dec 15;4(4):102560. doi: 10.1016/j.xpro.2023.102560. Epub 2023 Sep 23.

DOI:10.1016/j.xpro.2023.102560

PMID:37742174

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10522993/

Abstract

Mitochondrial respirometry allows for the comprehensive study of oxygen consumption within the electron transport system in tissues. However, limited techniques exist for analyzing frozen or biobanked intestinal tissues. Here, we present a protocol to evaluate the respiratory function of mitochondria in colonic tissues after cryopreservation at -80°C. We describe steps for rat dissection, respirometry calibration, and tissue preparation. We then detail measurement of oxygen respiration and protein concentration. This protocol facilitates the retrospective analysis of mitochondrial respiration in frozen tissue.

摘要

线粒体呼吸测量法允许全面研究组织中电子传递系统内的耗氧量。然而，用于分析冷冻或生物样本库肠组织的技术有限。在此，我们提出了一种在 -80°C 冷冻保存后评估结肠组织中线粒体呼吸功能的方案。我们描述了大鼠解剖、呼吸测量校准和组织准备的步骤。然后，我们详细说明了氧呼吸和蛋白质浓度的测量。该方案促进了对冷冻组织中线粒体呼吸的回顾性分析。