Oxygen-Independent Assays to Measure Mitochondrial Function in Mammals

Tenzin Tseyang; Jonathan Valeros; Paula Vo; Jessica B. Spinelli

doi:10.3791/65184

Journal

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Biochemistry

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Oxygen-Independent Assays to Measure Mitochondrial Function in Mammals

JoVE Journal
Biochemistry

This content is Free Access.

JoVE Journal Biochemistry

Oxygen-Independent Assays to Measure Mitochondrial Function in Mammals

2,394 Views

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05:59 min

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May 19, 2023

DOI:

10.3791/65184-v

DOI:

10.3791/65184-v

•

05:59 min

May 19, 2023

•

43 Views

Tenzin Tseyang*¹, Jonathan Valeros*¹, Paula Vo*¹, Jessica B. Spinelli^1,2

¹Program in Molecular Medicine,University of Massachusetts Chan Medical School, ²Cancer Center,University of Massachusetts Chan Medical School

Transcript

Our research focuses on how mitochondria sense and adapt to metabolic stressors. One key metabolic stressor is hypoxia, which occurs when cells are starved of oxygen. We are developing better assays to study mitochondrial function in low-oxygen environments.

Higher resolution mass spectrometry was employed to study cell and tissue metabolite levels. We used stable isotope tracing studies to track the fate of specific metabolites and uncover novel metabolic pathways relevant to healthy and diseased mammalian physiology. Group has found that mammalian mitochondria can sustain critical functions in low-oxygen environments.

Mechanistically, they employ fumarate as an alternative electronic scepter for the electron transport chain. This protocol compiles several assays that researchers can use to better assess mitochondrial function in mammalian cells. Importantly, these protocols can be used to study mitochondrial function in hypoxia, as classical respirometry experiments will not be sufficient.

Summary

Here, we present a compilation of assays to directly measure mitochondrial function in mammalian cells independently of their ability to consume molecular oxygen.