Isolation of Mitochondria From Fresh Mice Lung Tissue

Dayene de Assis Fernandes Caldeira; Dahienne Ferreira de Oliveira; João Paulo Cavalcanti-de-Albuquerque; Jose Hamilton Matheus Nascimento; Walter Araujo Zin; Leonardo Maciel

doi:10.3389/fphys.2021.748261

Isolation of Mitochondria From Fresh Mice Lung Tissue

Front Physiol. 2021 Nov 30:12:748261. doi: 10.3389/fphys.2021.748261. eCollection 2021.

Authors

Dayene de Assis Fernandes Caldeira¹, Dahienne Ferreira de Oliveira², João Paulo Cavalcanti-de-Albuquerque¹, Jose Hamilton Matheus Nascimento¹, Walter Araujo Zin¹, Leonardo Maciel^{1

3}

Affiliations

¹ Carlos Chagas Filho Institute of Biophysics, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
² Institute of Medical Biochemistry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
³ Professor Geraldo Cidade Campus, Federal University of Rio de Janeiro, Duque de Caxias, Brazil.

Abstract

Direct analysis of isolated mitochondria enables a better understanding of lung dysfunction. Despite well-defined mitochondrial isolation protocols applicable to other tissues, such as the brain, kidney, heart, and liver, a robust and reproductive protocol has not yet been advanced for the lung. We describe a protocol for the isolation of mitochondria from lung tissue aiming for functional analyses of mitochondrial O₂ consumption, transmembrane potential, reactive oxygen species (ROS) formation, ATP production, and swelling. We compared our protocol to that used for heart mitochondrial function that is well-established in the literature, and achieved similar results.

Keywords: ATP; O2-consumption; ROS; lung mitochondria isolation; mitochondrial assessment.