Comprehensive assessment of mitochondrial respiratory function in freshly isolated nephron segments

Allison McCrimmon, Mark Domondon, Regina F. Sultanova, Daria V. Ilatovskaya, Krisztian Stadler

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Changes in mitochondrial function are central to many forms of kidney disease, including acute injury, diabetic nephropathy, hypertension, and chronic kidney diseases. As such, there is an increasing need for reliable and fast methods for assessing mitochondrial respiratory function in renal cells. Despite being indispensable for many mechanistic studies, cultured cells or isolated mitochondria, however, often do not recapitulate in vivo or close to in vivo situations. Cultured and/or immortalized cells often change their bioenergetic profile and phenotype compared with in vivo or ex vivo situations, and isolated mitochondria are simply removed from their cellular milieu. This is especially important for extremely complex organs such as the kidney. Here, we report the development and validation of a new approach for the rapid assessment of mitochondrial oxygen consumption on freshly isolated glomeruli or proximal tubular fragments using Agilent SeaHorse XFe24 and XF96 Extracellular Flux Analyzers. We validated the technique in several healthy and diseased rodent models: the C57BL/6J mouse, the diabetic db/db mouse and matching db/+ control mouse, and the Dahl salt-sensitive rat. We compared the data to respiration from isolated mitochondria. The method can be adapted and used for the rapid assessment of mitochondrial oxygen consumption from any rodent model of the investigator's choice. The isolation methods presented here ensure viable and functional proximal tubular fragments and glomeruli, with a preserved cellular environment for studying mitochondrial function within the context of their surroundings and interactions.

Original languageEnglish (US)
Pages (from-to)F1237-F1245
JournalAmerican Journal of Physiology - Renal Physiology
Volume318
Issue number5
DOIs
StatePublished - May 2020
Externally publishedYes

Keywords

  • Bioenergetics
  • Glomeruli
  • Mitochondria
  • Proximal tubules

ASJC Scopus subject areas

  • Physiology
  • Urology

Fingerprint

Dive into the research topics of 'Comprehensive assessment of mitochondrial respiratory function in freshly isolated nephron segments'. Together they form a unique fingerprint.

Cite this