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A Platform for Mitochondrial Profiling in Enriched Kidney Segments Under Thermodynamic Control.

Stephen T Decker1, Precious C Opurum1,2, Ran Hee Choi1,2

  • 1Diabetes and Metabolism Research Center, University of Utah, Salt Lake City, Utah.

Biorxiv : the Preprint Server for Biology
|July 14, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a new method to measure kidney cell energy production in specific nephron segments. It reveals distinct mitochondrial functions across segments and identifies proximal tubule dysfunction in kidney disease models.

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Area of Science:

  • * Cellular and Molecular Physiology
  • * Renal Physiology
  • * Mitochondrial Biology

Background:

  • * Mitochondrial function is critical for kidney health but varies significantly across nephron segments.
  • * Existing methods lack the resolution to study cell-type-specific mitochondrial bioenergetics in situ.
  • * Understanding segment-specific bioenergetics is crucial for diagnosing and treating kidney diseases.

Purpose of the Study:

  • * To develop and validate a novel methodological platform for segment-resolved profiling of mitochondrial function in mouse nephron segments.
  • * To investigate the bioenergetic differences between various kidney nephron segments.
  • * To assess mitochondrial dysfunction in a model of adenine-induced nephropathy at a segment-specific level.

Main Methods:

  • * Mechanical sieving and adhesion-based enrichment for isolating specific nephron segments.
  • * Permeabilized high-resolution respirometry adapted with a creatine kinase clamp.
  • * Quantification of oxygen flux and mitochondrial membrane potential across defined free energies.

Main Results:

  • * Proximal tubules display high respiratory conductance and dynamic mitochondrial polarization.
  • * Distal tubules and glomeruli exhibit static membrane potential and low respiratory conductance.
  • * Adenine-induced nephropathy selectively impaired proximal tubule mitochondrial respiration and ATP production, undetectable in bulk isolates.

Conclusions:

  • * The developed platform enables precise, segment-resolved analysis of mitochondrial bioenergetics in native kidney tissue.
  • * Significant functional heterogeneity exists in mitochondrial bioenergetics across kidney nephron segments.
  • * This approach reveals segment-specific mitochondrial pathology in kidney disease, offering new diagnostic and therapeutic insights.