Cells in culture from rabbit medullary thick ascending limb of Henle's loop

Am J Physiol. 1989 May;256(5 Pt 1):C1070-81. doi: 10.1152/ajpcell.1989.256.5.C1070.

Abstract

Freshly isolated cells obtained from the medullary segment of the rabbit thick ascending limb of Henle's loop (mTALH) metabolize arachidonic acid (AA) primarily by the cytochrome P-450 monooxygenase pathway forming several products; a vasorelaxant and an inhibitor of Na+-K+-ATPase have been identified. These studies have been extended to mTALH cells in culture. The ability of cells isolated from 1-mo-old rabbits to grow in culture far surpassed that of cells isolated from adult rabbits, whereas similar cytochrome P-450-dependent AA metabolites were produced by freshly isolated cells from rabbits of both ages. Three-week-old mTALH cultures formed ouabain-sensitive "domes" when grown on plastic surfaces and developed transepithelial voltages (4.7 + 1.2 mV, n = 6) when grown on gas-permeable surfaces. Electron microscopy of the cells showed typical mTALH cell characteristics. The presence of Tamm-Horsfall protein, a surface membrane protein of mTALH cells, in 90-95% of the cells confirmed the homogeneity of the cultures. Although several environmental manipulations were tested, mTALH cells in culture did not produce the same cytochrome P-450-dependent AA metabolites as those produced by mTALH cells before culture. However, a cytochrome P-450-dependent AA metabolite that differs from the AA metabolites formed by freshly isolated mTALH cells was produced by hemin-treated mTALH and heterogenous cell cultures.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Arachidonic Acid
  • Arachidonic Acids / metabolism
  • Biological Transport
  • Cell Communication
  • Cells, Cultured
  • Cytochrome P-450 Enzyme System / analysis
  • Kidney Medulla / cytology*
  • Kidney Tubules / cytology*
  • Loop of Henle / cytology*
  • Loop of Henle / metabolism
  • Loop of Henle / ultrastructure
  • Male
  • Microscopy, Electron
  • Mucoproteins / analysis
  • NADP / metabolism
  • Rabbits
  • Sodium-Potassium-Exchanging ATPase / physiology
  • Uromodulin

Substances

  • Arachidonic Acids
  • Mucoproteins
  • Uromodulin
  • Arachidonic Acid
  • NADP
  • Cytochrome P-450 Enzyme System
  • Sodium-Potassium-Exchanging ATPase