AVP stimulates Na+ transport in primary cultures of rabbit cortical collecting duct cells.
Immunodissected rabbit cortical collecting duct (CCD) cells were grown in primary culture on permeable membrane supports. Transepithelial voltage, Na+, K+, and H+ gradients developed as expected for a mixed population of principal and intercalated cells. The amiloride-sensitive short-circuit current (Isc) was measured in Ussing chambers as an index of Na+ transport via apical membrane Na+ channels. Treatment of the cells in culture with 10 nM aldosterone for 48 h increased Isc from 7.4 +/- 1.4 to 19.3 +/- 3.2 microA/cm2. In contrast to the native rabbit CCD, 220 pM arginine vasopressin (AVP) produced a rapid and stable (greater than 60 min) increase in Isc to 15.8 +/- 2.0 and 29.0 +/- 3.8 microA/cm2 in untreated and aldosterone-treated cultures, respectively. Although prostaglandin E2 (PGE2) inhibits Na+ transport in the native rabbit CCD, it did not in the cultured cells, and it has previously been shown that PGE2 inhibition of AVP-dependent adenosine 3',5'-cyclic monophosphate production is lost in culture (W. K. Sonnenburg and W. L. Smith, J. Biol. Chem. 263: 6155-6160, 1988). We conclude that the development of a stable stimulation of Na+ transport by AVP is linked to the loss of the inhibitory effects of PGE2.
Canessa
C M
CM
Department of Physiology and Biophysics, University of Alabama, Birmingham 35294.
Schafer
J A
JA
D000450:Aldosterone / Q000494:pharmacology; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal; D001127:Arginine Vasopressin / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D004848:Epithelium / Q000187:drug effects / Q000502:physiology; D005665:Furosemide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007672:Kidney Cortex / Q000187:drug effects / Q000502:physiology*; D007685:Kidney Tubules, Collecting / Q000187:drug effects / Q000502:physiology*; D007700:Kinetics; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism; D011817:Rabbits; D012964:Sodium / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D001127:Arginine Vasopressin; D000450:Aldosterone; D010042:Ouabain; D000584:Amiloride; D005665:Furosemide; D012964:Sodium; D011188:Potassium