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Super-resolution microscopy of mitochondrial mRNAs.

Stefan Stoldt1,2,3, Frederike Maass1,2,3, Michael Weber4

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Researchers visualized individual mitochondrial messenger RNA (mRNA) molecules using super-resolution microscopy. This technique reveals how mRNA distribution and shape change in cells, offering new insights into mitochondrial gene regulation.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Mitochondria possess their own DNA (mtDNA) and gene expression machinery.
  • The spatial distribution of mitochondrial mRNA and proteins is poorly understood due to microscopy limitations.

Purpose of the Study:

  • To develop and apply super-resolution microscopy techniques to visualize individual mitochondrial mRNA molecules and associated proteins.
  • To investigate the spatial organization and dynamics of mitochondrial mRNA in response to cellular stress and disease.

Main Methods:

  • Single-molecule fluorescence in situ hybridization (smFISH) combined with STED and MINFLUX super-resolution microscopy.
  • Visualization of individual mitochondrial mRNA molecules and RNA-binding proteins.
  • Analysis of mRNA distribution, shape, and proximity to ribosomes in mammalian cells.

Main Results:

  • STED nanoscopy revealed spatial relationships between different mRNA species and proteins like GRSF1.
  • Adaptive changes in mRNA distribution and quantity were observed in stressed cells and patient-derived cell lines.
  • STED-smFISH showed mRNA release during apoptosis, and MINFLUX revealed mRNA folding and proximity to mitochondrial ribosomes.

Conclusions:

  • Super-resolution microscopy enables visualization of mitochondrial mRNA and associated proteins at the single-molecule level.
  • Mitochondrial mRNA distribution and organization are dynamic and adapt to cellular conditions, including apoptosis.
  • These advanced imaging techniques provide new avenues for studying mitochondrial gene regulation in health and disease.