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Am J Physiol Regul Integr Comp Physiol 277: R1725-R1732, 1999;
0363-6119/99 $5.00
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Vol. 277, Issue 6, R1725-R1732, December 1999

Mode of activation of salt secretion by C-type natriuretic peptide in the shark rectal gland

Patricio Silva1,2, Richard J. Solomon1,3,4, and Franklin H. Epstein1,3

2 Department of Medicine, Temple University Hospital, Philadelphia, Pennsylvania 19140; 3 Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston 02215; 4 Joslin Diabetes Center, Boston, Massachusetts 02215; and 1 The Mount Desert Island Biological Laboratory, Salsbury Cove, Maine 04672

We studied the modes of activation of the salt-secreting rectal gland of the spiny dogfish, Squalus acanthias, by the native cardiac peptide CNP. The stimulatory action of CNP in isolated perfused glands is inhibited by 10 mM procaine, presumably by blocking release of vasoactive intestinal peptide (VIP) from nerves. Procaine reduces the slope of the dose-response curve of human CNP and that of shark CNP (each P < 0.0001). CNP increases short-circuit current in cultured rectal gland cells from 4.8 ± 1.6 to 27.0 ± 7.8 µA/cm2. It also stimulates the secretion of chloride in isolated perfused glands in the presence of 10 mM procaine from 72 ± 31 to 652 ± 173 µeq · h-1 · g-1. These results suggest that CNP has a direct cellular action not mediated by the neural release of VIP. The residual stimulation of perfused glands in the presence of procaine was almost completely inhibited by staurosporine [10 nM; an inhibitor of protein kinase C (PKC)] from 652 ± 173 to 237 ± 61 µeq · h-1 · g-1. Although CNP stimulates guanylyl cyclase in shark rectal gland, chloride secretion of perfused glands was not elicited by 8-bromoadenosine-cGMP (8-BrcGMP) alone nor by the activator of PKC phorbol ester. The combination of PKC activation and 8-BrcGMP infusion, however, stimulated chloride secretion in perfused glands from 94 ± 30 to 506 ± 61 µeq · h-1 · g-1, a level comparable to that observed in glands blocked with procaine. Several parallel pathways appear to be synergistic in activating chloride secretion stimulated by CNP in the rectal gland.

chloride secretion; guanylyl cyclase; guanosine 3',5'-cyclic monophosphate; protein kinase C; phorbol esters


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