Role of F-actin in the activation of Na(+)-K(+)-Cl- cotransport by forskolin and vasopressin in mouse kidney cultured thick ascending limb cells.
The influence of microtubules and F-actin on Na(+)-K(+)-Cl- cotransport was investigated in cultured cells derived from outer-medullary thick ascending limb tubules microdissected from the mouse kidney. The cultured cells contained Tamm-Horsfall protein, produced cAMP in response to dD-arginine vasopressin (dD-AVP), isoproterenol, prostaglandin E2 and forskolin (FK), and exhibited an ouabain-resistant furosemide-sensitive (Or-Fs) component of 86Rb+ influx mediated by the Na(+)-K(+)-Cl- cotransporter. Both FK and dD-AVP stimulated the Or-Fs component of Rb+ influx. Neither agent altered the tubulin and cytokeratin networks nor the shape of the tight junction using a specific anti-ZO-1 antibody. In contrast, they did induce a marked redistribution of F-actin to the periphery of the cells delineating the tight junctions. Preincubation of the cells with nocodazole, to disrupt microtubules, did not alter the FK- or dD-AVP-elicited Or-Fs Rb+ influx. In contrast, phalloidin and NBD-phallicidin, which stabilize F-actin, markedly impaired the stimulation of Na(+)-K(+)-Cl- cotransport by FK or dD-AVP, without affecting the Na(+)-K+ ATPase pumps and the rate constant of 36Cl- and 86Rb+ efflux. These results strongly suggested that cAMP-stimulated Na(+)-K(+)-Cl- cotransport is linked to F-actin in renal TAL cells.
Wu
M S
MS
Institut National de la Santé et de la Recherche Médicale (INSERM), Unité 246, Institut Fédératif de Recherche Xavier Bichat, Paris, France.
Bens
M
M
Cluzeaud
F
F
Vandewalle
A
A
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001127:Arginine Vasopressin / Q000494:pharmacology*; D002478:Cells, Cultured; D002712:Chlorides / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology*; D000242:Cyclic AMP / Q000378:metabolism; D003599:Cytoskeleton / Q000187:drug effects; D015232:Dinoprostone / Q000494:pharmacology; D004232:Diuretics / Q000494:pharmacology; D017136:Ion Transport / Q000502:physiology*; D007545:Isoproterenol / Q000494:pharmacology; D008138:Loop of Henle / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051379:Mice; D015739:Nocodazole / Q000494:pharmacology; D010042:Ouabain / Q000494:pharmacology; D010281:Parathyroid Hormone / Q000494:pharmacology; D010590:Phalloidine / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism
D000199:Actins; D002712:Chlorides; D004232:Diuretics; D010281:Parathyroid Hormone; D001127:Arginine Vasopressin; D010590:Phalloidine; D005576:Colforsin; D010042:Ouabain; D012964:Sodium; D000242:Cyclic AMP; D015232:Dinoprostone; D007545:Isoproterenol; D011188:Potassium; D015739:Nocodazole; D015056:1-Methyl-3-isobutylxanthine