Bioenergetic characterization of mouse podocytes.
نویسندگان
چکیده
Mitochondrial dysfunction contributes to podocyte injury, but normal podocyte bioenergetics have not been characterized. We measured oxygen consumption rates (OCR) and extracellular acidification rates (ECAR), using a transformed mouse podocyte cell line and the Seahorse Bioscience XF24 Extracellular Flux Analyzer. Basal OCR and ECAR were 55.2 +/- 9.9 pmol/min and 3.1 +/- 1.9 milli-pH units/min, respectively. The complex V inhibitor oligomycin reduced OCR to approximately 45% of baseline rates, indicating that approximately 55% of cellular oxygen consumption was coupled to ATP synthesis. Rotenone, a complex I inhibitor, reduced OCR to approximately 25% of the baseline rates, suggesting that mitochondrial respiration accounted for approximately 75% of the total cellular respiration. Thus approximately 75% of mitochondrial respiration was coupled to ATP synthesis and approximately 25% was accounted for by proton leak. Carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), which uncouples electron transport from ATP generation, increased OCR and ECAR to approximately 360% and 840% of control levels. FCCP plus rotenone reduced ATP content by 60%, the glycolysis inhibitor 2-deoxyglucose reduced ATP by 35%, and 2-deoxyglucose in combination with FCCP or rotenone reduced ATP by >85%. The lactate dehydrogenase inhibitor oxamate and 2-deoxyglucose did not reduce ECAR, and 2-deoxyglucose had no effect on OCR, although 2-deoxyglucose reduced ATP content by 25%. Mitochondrial uncoupling induced by FCCP was associated with increased OCR with certain substrates, including lactate, glucose, pyruvate, and palmitate. Replication of these experiments in primary mouse podocytes yielded similar data. We conclude that mitochondria play the primary role in maintaining podocyte energy homeostasis, while glycolysis makes a lesser contribution.
منابع مشابه
Large Deformation Characterization of Mouse Oocyte Cell Under Needle Injection Experiment
In order to better understand the mechanical properties of biological cells, characterization and investigation of their material behavior is necessary. In this paper hyperelastic Neo-Hookean material is used to characterize the mechanical properties of mouse oocyte cell. It has been assumed that the cell behaves as continuous, isotropic, nonlinear and homogenous material for modeling. Then, by...
متن کاملStatin-sensitive endocytosis of albumin by glomerular podocytes.
Glomerular podocytes are critical regulators of glomerular permeability via the slit diaphragm and may play a role in cleaning the glomerular filter. Whether podocytes are able to endocytose proteins is uncertain. We studied protein endocytosis in conditionally immortalized mouse and human podocytes using FITC-albumin by direct quantitative assay and by fluorescence microscopy and electron micr...
متن کاملCharacterization of renin-angiotensin system enzyme activities in cultured mouse podocytes
Velez JC, Bland AM, Arthur JM, Raymond JR, Janech MG. Characterization of renin-angiotensin system enzyme activities in cultured mouse podocytes. Am J Physiol Renal Physiol 293: F398–F407, 2007. First published April 7, 2007; doi:10.1152/ajprenal.00050.2007.—Intraglomerular ANG II has been linked to glomerular injury. However, little is known about the contribution of podocytes (POD) to intragl...
متن کاملTetracycline-inducible gene expression in conditionally immortalized mouse podocytes.
BACKGROUND Conditionally immortalized podocytes are valuable research tools but are difficult to efficiently transfect and do not provide graded transgene expression. METHODS Conditionally immortalized mouse podocyte cell lines were established employing a tetracycline-inducible system. Glomerular cells, isolated from transgenic mice bear- ing two transgenes, NPHS2-reverse tetracycline-contro...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- American journal of physiology. Cell physiology
دوره 299 2 شماره
صفحات -
تاریخ انتشار 2010