Osmoregulation of GPC:choline phosphodiesterase in MDCK cells: Different effects of urea and NaCl

E. D. Kwon, K. Zablocki, Yong Jung Kyu Yong Jung, E. M. Peters, A. Garcia-Perez, M. B. Burg

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

organic osmolyte, glycerophosphocholine (GPC), accumulates in renal cells in response to high concentrations of either NaCl or urea, despite the very different effects of these solutes on cell function and volume. Together, high levels of these solutes increase GPC amount in Madin-Darby canine kidney cells by inhibiting its enzymatic degradation. The present study tests the effects of NaCl and urea, individually, on GPC accumulation and its degradation. A technique was developed to determine the absolute rate of GPC degradation by measuring the initial rate of disappearance of [3H]GPC (pulsed into the cells by hypotonic shock) and the specific activity of GPC in the cells. The mass of GPC in the cells was measured by another newly developed method, a sensitive chemiluminescent assay. We find that exposure to high NaCl or urea decreases the absolute rate of cellular GPC degradation by approximately one-half during the first 20.5 h. Reductions in GPC degradation are accompanied by commensurate decreases in the activity of GPC:choline phosphodiesterase (GPC:PDE; EC 3.1.4.2), an enzyme that catalyzes degradation of GPC. Activity of GPC:PDE falls >50% in cells exposed for 2 h to high osmolality. Inhibition is sustained for 7 days with high urea alone. In contrast, with high NaCl alone, GPC:PDE activity reverts to control values by 7 days, by which time synthesis of GPC is increased, accounting for sustained GPC accumulation. Collectively, these data suggest that GPC accumulation in response to either high NaCl or urea occurs initially by inhibition of its degradation but that the effect of NaCl on degradation differs, in that it is transient, while that of urea is sustained.

Original languageEnglish (US)
Pages (from-to)C35-C41
JournalAmerican Journal of Physiology - Cell Physiology
Volume269
Issue number1 38-1
DOIs
StatePublished - 1995

Keywords

  • Madin-Darby canine kidney cells
  • glycerophosphocholine
  • hyperosmotic stress
  • methylamines
  • organic osmolytes
  • renal medulla

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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