title: Cells in culture from rabbit medullary thick ascending limb of Henle's loop.
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.
authors.affiliation: Department of Pharmacology, New York Medical College, Valhalla 10595.
authors.forename: E D
authors.initials: ED
authors.lastname: Drugge
authors.forename: M A
authors.initials: MA
authors.lastname: Carroll
authors.forename: J C
authors.initials: JC
authors.lastname: McGiff
mesh_terms: D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D001692:Biological Transport; D002450:Cell Communication; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D007679:Kidney Medulla / Q000166:cytology*; D007684:Kidney Tubules / Q000166:cytology*; D008138:Loop of Henle / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D009088:Mucoproteins / Q000032:analysis; D009249:NADP / Q000378:metabolism; D011817:Rabbits; D000254:Sodium-Potassium-Exchanging ATPase / Q000502:physiology; D058949:Uromodulin
chemical_list: D001095:Arachidonic Acids; D009088:Mucoproteins; D058949:Uromodulin; D016718:Arachidonic Acid; D009249:NADP; D003577:Cytochrome P-450 Enzyme System; D000254:Sodium-Potassium-Exchanging ATPase