Triglyceride depletion of brown adipose tissue enables analysis of mitochondrial respiratory function in permeabilized biopsies

Autor(es)

Dechandt, Carlos R. P. ; Couto-Lima, Carlos A. ; Alberici, Luciane C.

• Tipo de documento: Artigo Científico
• Fonte: Analytical Biochemistry; v. 515, p. 55-60, DEC 15 2016.
• Citações Web of Science: 3

Resumo

The research on mitochondrial functions in adipocytes has increasingly evidenced that mitochondria plays an important role in the onset and/or progression of obesity and related pathologies. Mitochondrial function in brown adipose tissue (BAT) has been classically assessed by measuring either the levels/activity of mitochondrial enzymes, or the respiration in isolated mitochondria. Isolation of mitochondria is not advantageous because it demands significant time and amount of tissue and, as tissue homogenates, disrupts biochemical and physical connections of mitochondria within the cell. Here, we described a new and efficient protocol to analyze the mitochondrial respiratory states in BAT biopsies that relies on intracellular triglyceride depletion followed by tissue permeabilization. In addition to minimizing tissue requirements to similar to 17 mg wet weight, the proposed protocol enabled analysis of all mitochondrial respiratory states, including phosphorylation (OXPHOS), no-phosphorylation (LEAK), and uncoupled (ETS) states, as well as the use of substrates for complex I, complex II, and cytochrome c; together, these features demonstrated mitochondrial integrity and validated the preparation efficacy. Therefore, the protocol described here increases the possibilities of answering physiological questions related to small BAT regions of human and animal models, which shall help to unravel the mechanisms that regulate mitochondrial function in health and disease. (C) 2016 Elsevier Inc. All rights reserved. (AU)

• Processo FAPESP: 10/17259-9 - Estudos sobre mecanismos de desacoplamento mitocondriais por ácidos graxos não-esterificados como estratégia de prevenção/tratamento da obesidade
• Beneficiário: Luciane Carla Alberici
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores