AJP - Cell Physiology, Vol 257, Issue 2 C333-C340, Copyright © 1989 by American Physiological Society

ARTICLES

Glycolytic and oxidative metabolism in primary renal proximal tubule cultures

K. G. Dickman and L. J. Mandel
Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710.

Cultured cells often exhibit alterations in energy metabolism (increased glycolytic activity and decreased oxidative metabolism) during adaptation to the culture environment. The role of hypoxia as a mediator of these effects was examined by comparison of metabolism in primary rabbit renal proximal tubule (RPT) cultures maintained in stationary culture dishes (DISH), shaking Erlenmeyer flasks (SHAKE), and DISH cultures transferred back to SHAKE conditions (RESHAKE). Both oxidative metabolism and transport capacity were fully preserved in SHAKE cultures over a 24-h period. In contrast, within 6 h, DISH cultures exhibited a continuous decline in transport-dependent and -independent oxygen consumption, respiratory capacity, and ATP and K+ contents. The loss of oxidative metabolism in DISH cultures was accompanied by stimulation of lactate production, detectable within 1 h after plating. Comparison of metabolic properties of DISH cultures to those of RPT exposed to graded levels of hypoxia suggested that medium oxygen tensions may be as low as 1-3% in DISH cultures. RESHAKE cultures exhibited metabolic properties comparable to those of SHAKE cultures, indicating reversibility of DISH culture metabolism on reoxygenation. We concluded that DISH cultures rapidly become hypoxic as a consequence of static culture conditions. Shaking suspension cultures may provide a more metabolically appropriate model for long-term in vitro studies.

This article has been cited by other articles:

				A. Deng, C. M. Miracle, M. Lortie, J. Satriano, F. B. Gabbai, K. A. Munger, S. C. Thomson, and R. C. Blantz Kidney oxygen consumption, carbonic anhydrase, and proton secretion Am J Physiol Renal Physiol, May 1, 2006; 290(5): F1009 - F1015. [Abstract] [Full Text] [PDF]

				Z. Bebok, A. Tousson, L. M. Schwiebert, and C. J. Venglarik Improved oxygenation promotes CFTR maturation and trafficking in MDCK monolayers Am J Physiol Cell Physiol, January 1, 2001; 280(1): C135 - C145. [Abstract] [Full Text] [PDF]

				P. White, R. B. Doctor, R. H. Dahl, and J. Chen Coincident microvillar actin bundle disruption and perinuclear actin sequestration in anoxic proximal tubule Am J Physiol Renal Physiol, June 1, 2000; 278(6): F886 - F893. [Abstract] [Full Text] [PDF]

				C. Bolon, C. Gauthier, and H. Simonnet Glycolysis inhibition by palmitate in renal cells cultured in a two-chamber system Am J Physiol Cell Physiol, November 1, 1997; 273(5): C1732 - C1738. [Abstract] [Full Text] [PDF]

				J. Chen and L. J. Mandel Role of water and electrolyte influxes in anoxic plasma membrane disruption Am J Physiol Cell Physiol, October 1, 1997; 273(4): C1341 - C1348. [Abstract] [Full Text] [PDF]

				C. A. Blaze, P. J. Mannon, S. R. Vigna, A. R. Kherani, and B. A. Benjamin Peptide YY receptor distribution and subtype in the kidney: effect on renal hemodynamics and function in rats Am J Physiol Renal Physiol, October 1, 1997; 273(4): F545 - F553. [Abstract] [Full Text] [PDF]

				S. P. Soltoff Rottlerin Is a Mitochondrial Uncoupler That Decreases Cellular ATP Levels and Indirectly Blocks Protein Kinase Cdelta Tyrosine Phosphorylation J. Biol. Chem., October 5, 2001; 276(41): 37986 - 37992. [Abstract] [Full Text] [PDF]