MitoRACE: evaluating mitochondrial function in vivo and in single cells with subcellular resolution using multiphoton...

Area of Science:

• Cell Biology
• Metabolic Imaging
• Mitochondrial Function

Background:

• Mitochondria are crucial in various cell types and diseases.
• Existing methods lack the resolution to assess mitochondrial heterogeneity.
• Direct in vivo assessment of mitochondrial function is limited.

Purpose of the Study:

• To introduce a novel metabolic imaging approach for direct measurement of mitochondrial energy conversion.
• To achieve single-cell and subcellular resolution in assessing mitochondrial function.
• To provide a platform for evaluating mitochondrial function in vivo and in cell culture.

Main Methods:

• Developed the mitochondrial redox after cyanide experiment (mitoRACE).
• Utilized NADH autofluorescence kinetics to measure mitochondrial NADH flux.
• Assessed sensitivity to physiological and pharmacological perturbations in vivo.

Main Results:

• mitoRACE provides direct measures of mitochondrial energy conversion with single-cell and subcellular resolution.
• The technique is sensitive to physiological and pharmacological changes in mitochondrial flux.
• Demonstrated higher energetic flux in neuronal neurites compared to cell bodies.

Conclusions:

• Metabolic imaging with mitoRACE is a highly adaptable platform for evaluating mitochondrial function.
• mitoRACE offers potential for broad applications in energy metabolism research.
• The technique enables detailed analysis of mitochondrial function heterogeneity.