Acute exposure to C60 fullerene damages pulmonary mitochondrial function and mechanics

Mitochondria in Focus: From Function to Therapeutic Strategies in Chronic Lung Diseases

“…Mitochondrial ROS (mtROS) can act as a second messenger, promoting physiologic signals of cellular stress and leading to mitochondrial translocation ( 64 , 67 ). Particularly in the lungs, due to their anatomy and continuous exposure to the environment, mitochondrial respiration is an important endogenous source of oxidative stress ( 71 , 72 ). When in excess, mtROS leads to uncoupled ETC, calcium imbalance, impaired communication between the ER and mitochondria, and damaged mtDNA, and acts as an inflammatory signal ( 73 ).…”

Isolation of Mitochondria From Fresh Mice Lung Tissue

“…We used herein a new protocol with specific and detailed steps aiming to improve mitochondrial isolation from lung tissue. It was abridgedly published ( Caldeira et al, 2021 ) but not tested against a well-documented and broadly used one ( Gedik et al, 2017 ). This new protocol improves the acquisition of a robust and preserved sample of isolated mitochondria, allowing a range of analyses with the same sample, increasing mitochondria viability and experimental reproducibility.…”

“…The assessment of mitochondrial function in organs and tissues is essential for a better understanding of their biochemistry, physiology, and pathophysiology ( Weissig, 2005 ; Picard et al, 2011 ; Meyer et al, 2017 ; Murphy and Hartley, 2018 ). The evaluation of mitochondrial function is usually accomplished in isolated mitochondria ( Picard et al, 2011 ; Gedik et al, 2017 ; Maciel et al, 2020 , 2021 ; Caldeira et al, 2021 ) permeabilized fibers ( Perry et al, 2013 ) or cells ( Perry et al, 2013 ). These techniques are very well-defined in several types of tissue, e.g., heart ( Gedik et al, 2017 ), kidney ( Schulz et al, 2015 ), liver ( Goudarzi et al, 2018 ), adipose tissue ( Matta et al, 2021 ), and brain ( Marques Neto et al, 2020 ), presenting peculiarities and different indications depending on the tissue and the purpose of the investigation.…”

“…The methodological difficulty of obtaining viable lung mitochondria derives mainly from an elevated fatty acid content, low amount of mitochondria in the cell, fibrous and air-filled tissue, and the required amount of tissue ( Spear and Lumeng, 1978 ; Kuznetsov et al, 2008 ; Lanza and Sreekumaran Nair, 2009 ). Recently, we have shown that the isolation of pulmonary mitochondria with preserved structure and function is possible by means of adaptations of existing techniques and standardization of a specific method of isolation by differentiated centrifugation ( Caldeira et al, 2021 ). However, the differences related to mitochondrial isolation procedures for obtaining these well-preserved pulmonary mitochondria have not been addressed in detail, and the characteristics of reagents and equipment have not been fully described ( Caldeira et al, 2021 ).…”

“…Recently, we have shown that the isolation of pulmonary mitochondria with preserved structure and function is possible by means of adaptations of existing techniques and standardization of a specific method of isolation by differentiated centrifugation ( Caldeira et al, 2021 ). However, the differences related to mitochondrial isolation procedures for obtaining these well-preserved pulmonary mitochondria have not been addressed in detail, and the characteristics of reagents and equipment have not been fully described ( Caldeira et al, 2021 ). Therefore, the main objective of this article is to provide a practical step-by-step user protocol upgraded to isolate pulmonary mitochondria.…”