Long-term effects of vasopressin on the subcellular localization of ENaC in the renal collecting system.
Previous studies revealed that chronic (days) vasopressin treatment stimulates amiloride-sensitive sodium transport in isolated renal cortical collecting ducts and increases the abundance of beta- and gamma-subunits of the epithelial sodium channel (ENaC) in the kidney. The aim of the present work was to investigate in vivo the cellular basis of these effects. The long-term effect of V2 vasopressin agonist (1-deamino-8-D-arginine vasopressin (dDAVP)) on the abundance and subcellular localization of ENaC along the rat renal collecting system was determined by immunohistochemistry and laser confocal microscopy. Moreover, we studied by real-time reverse transcriptase-polymerase chain reaction the effect of vasopressin on proteins implicated in the regulation of ENaC (Nedd4-2, prostasin, Sgk1). After 5 days of administration, dDAVP markedly increased the intracellular pool of the beta- and gamma-ENaC subunits in the principal cells, with an increasing gradient from connecting tubule to the outer medullary collecting duct, but did not increase any subunit at the cell surface. The apical immunostaining of ENaC increased in response to sodium restriction, as expected, but dDAVP did not further enhance this apical labelling. dDAVP increased the gene expression of prostasin in the cortex but not that of Nedd4-2 and Sgk1. These findings suggest that the previously reported increase in sodium transport induced by sustained stimulation of vasopressin V2 receptor is probably mediated by other mechanism than an increase in the apical density of ENaC.
Sauter
D
D
Institute of Anatomy, University of Zürich, Zürich, Switzerland.
Fernandes
S
S
Goncalves-Mendes
N
N
Boulkroun
S
S
Bankir
L
L
Loffing
J
J
Bouby
N
N
D000450:Aldosterone / Q000032:analysis / Q000097:blood; D000818:Animals; D003894:Deamino Arginine Vasopressin / Q000494:pharmacology; D004326:Drinking; D053503:Epithelial Sodium Channels; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects; D007150:Immunohistochemistry; D007672:Kidney Cortex / Q000737:chemistry; D007685:Kidney Tubules, Collecting / Q000737:chemistry*; D008297:Male; D018613:Microscopy, Confocal; D051381:Rats; D011910:Rats, Brattleboro; D017207:Rats, Sprague-Dawley; D017483:Receptors, Vasopressin / Q000032:analysis / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000032:analysis / Q000235:genetics; D015222:Sodium Channels / Q000032:analysis*; D012982:Sodium, Dietary / Q000494:pharmacology; D013997:Time Factors; D014667:Vasopressins / Q000097:blood / Q000494:pharmacology*; D000099271:Prostasin
D000450:Aldosterone; D003894:Deamino Arginine Vasopressin; D053503:Epithelial Sodium Channels; D017483:Receptors, Vasopressin; D012697:Serine Endopeptidases; D015222:Sodium Channels; D012982:Sodium, Dietary; D014667:Vasopressins; D000099271:Prostasin