CRABS-ROC, A respirometry protocol for overcoming substrate limitations, reveals excess brain mitochondrial Complex I capacity.

ABSTRACT: Mitochondrial bioenergetic competency in cells is frequently assessed by the Mito Stress Test protocol, which includes uncoupler addition for evaluating respiratory capacity. The uncoupled oxygen consumption rate (OCR) is usually defined as maximal respiration, with little consideration of whether the measured rate is restricted by substrate supply. In this study, we show that the uncoupled OCR is substrate-limited in rat primary cortical neurons and isolated mouse forebrain synaptosomes. We use a different respirometry protocol we name CRABS-ROC (Complex Respirometry Assay Bypassing Substrate-Restricted Oxygen Consumption) that enables evaluation of individual electron transport chain (ETC) complex capacities using saturating levels of substrates to bypass this restriction. Optimization o

SUBMITTER: Zhang N 

PROVIDER: S-EPMC12776583 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature