Dahdah 2019 MiP2019

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Norma Dahdah
Approaches to assess mitochondrial respiration in white adipose tissue.

Link: MiP2019

Dahdah N, Gama-Perez P, Gonzalez-Franquesa A, Garcia-Roves PM (2019)

Event: MiP2019

COST Action MitoEAGLE

Obesity is currently a global health problem that has reached epidemic proportions, with the consequent increase in the prevalence of its comorbidities, such as type 2 diabetes (T2D), cardiovascular disease, various cancers… White adipose tissue (WAT) is a fundamental organ in the regulation of whole-body metabolism and glucose homeostasis through its energy storage capability and its adipokine-mediated endocrine function. An association between the impaired mitochondrial function in WAT and the deranged glucose homeostasis found in obesity has been uncovered [1]. Mitochondrial function has been assessed in isolated mitochondria, in isolated adipocytes or in homogenized tissue, depending on the purpose of the experiments. The different methods and protocols used for these assessments have strengths and weaknesses [2]. When measuring mitochondrial respiration, the disadvantage of using isolated mitochondria is the high amount of tissue needed to yield one sample and the loss of damaged mitochondria through the isolation process. With isolated adipocytes, the protocol is time consuming and might lead to the lysis and loss of some adipocytes. Adipose tissue homogenization and permeabilization requires mechanical and chemical disruptions. Moreover, there will be a release of fatty acids and/or triglycerides when permeabilizing the cells. In addition, normalization of the samples is different between protocols and between the groups adopting these protocols.

Our aim is to set the basis for an open discussion about these approaches in order to standardize the protocols for better reliability and reproducibility.

We initiated the first steps to address this issue by measuring mitochondrial respiration in mouse epididymal WAT homogenates and adipocytes (isolated from the same tissue) and by measuring mitochondrial respiration under different BSA concentrations in the respiration media (MiR05) in order to reduce the assumed fatty acids-derived lipotoxicity.

The conducted experiments are preliminary and not enough to draw conclusions. More action and participation are needed in order to standardize a protocol for assessing mitochondrial respiration in white adipose tissue.


Bioblast editor: Plangger M, Tindle-Solomon L O2k-Network Lab: ES Barcelona Garcia-Roves PM


Labels: MiParea: Respiration, Instruments;methods 


Organism: Mouse  Tissue;cell: Fat  Preparation: Homogenate, Isolated mitochondria, Intact cells 



HRR: Oxygraph-2k 


Affiliations

Dahdah N(1), Gama-Perez P(1), Gonzalez-Franquesa A(2), Garcia-Roves PM(1)
  1. Dept Physiological Sciences, Univ Barcelona, Spain
  2. Univ Copenhagen, Denmark. - [email protected]

References

  1. Schöttl T, Kappler L, Braun K, Fromme T, Klingenspor M (2015) Limited mitochondrial capacity of visceral versus subcutaneous white adipocytes in male C57BL/6N mice. Endocrinology 156:923-33.
  2. Brand MD, Nicholls DG (1993) Assessing mitochondrial dysfunction in cells. Biochem J 435:297-312.