Regulation of renal epithelial sodium channels.
The high selectivity, low conductance, amiloride-blockable, sodium channel of the mammalian distal nephron (i.e. cortical collecting tubule) is the site of discretionary regulation which allows maintainance of total body sodium balance. In order to understand the physiological events that participate in this regulation, we have used the patch-clamp technique which allows us to measure individual Na+ channel currents and permits access to the cytosolic side of the channel-protein as well as its associated regulatory components. Most of our experiments have utilized the A6 amphibian renal cell line, which when grown on permeable supports is an excellent model for the mammalian distal nephron. Different mechanisms have been examined: (1) regulation by hormonal factors such as Anti-Diuretic Hormone (ADH) and aldosterone, (2) regulation by G-proteins, (3) modulation by protein kinase C (PK-C), and (4) modulation by products of arachidonic acid metabolism. Consistent with noise analysis of tight epithelial tissues, ADH treatment increased the number of active channels in apical membrane patches of A6 cells, without any apparent change in the open probability (Po) of the individual channels. Agents that increased intracellular cAMP mimicked the effects of ADH. In contrast, aldosterone was found to act through a dramatic increase in Po rather than through changes in channel density. Inhibition of methylation by deazaadenosine antagonizes the stimulatory effect of aldosterone. In excised inside-out patches GTP gamma S inhibits channel activity, whereas GDP beta S or pertussis toxin stimulates activity suggesting regulatory control by G-proteins. PK-C has been shown to contribute to 'feed-back inhibition' of apical Na+ conductance in tight epithelia.(ABSTRACT TRUNCATED AT 250 WORDS)
Duchatelle
P
P
Department of Physiology, Emory University School of Medicine, Atlanta, Georgia 30322.
Ohara
A
A
Ling
B N
BN
Kemendy
A E
AE
Kokko
K E
KE
Matsumoto
P S
PS
Eaton
D C
DC
D000450:Aldosterone / Q000502:physiology; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000502:physiology; D003599:Cytoskeleton / Q000502:physiology; D015777:Eicosanoids / Q000502:physiology; D004848:Epithelium / Q000502:physiology; D019204:GTP-Binding Proteins / Q000502:physiology; D066298:In Vitro Techniques; D015640:Ion Channel Gating; D007668:Kidney / Q000502:physiology*; D008564:Membrane Potentials; D011494:Protein Kinases / Q000502:physiology; D011817:Rabbits; D051381:Rats; D015222:Sodium Channels / Q000502:physiology*; D014667:Vasopressins / Q000502:physiology
D015777:Eicosanoids; D015222:Sodium Channels; D014667:Vasopressins; D000450:Aldosterone; D000584:Amiloride; D011494:Protein Kinases; D019204:GTP-Binding Proteins; D002118:Calcium