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Am J Physiol Renal Physiol 271: F762-F769, 1996;
0363-6127/96 $5.00
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AJP - Renal Physiology, Vol 271, Issue 3 762-F769, Copyright © 1996 by American Physiological Society

ARTICLES

The beta 1- and beta 2-adrenoceptor subtypes in cultured rat inner medullary collecting duct cells

G. Yasuda, L. Sun, H. C. Lee, S. Umemura and W. B. Jeffries
Second Department of Internal Medicine, Yokohama City University School of Medicine, Japan.

We investigated beta-adrenoceptor subtype(s) expressed in cultured rat inner medullary collecting duct (IMCD) cells. In radioligand binding assay, [125I]iodocyanopindolol bound to IMCD cell membranes, representing a single class of binding sites (dissociation constant = 96.1 pM, maximum binding capacity = 18.2 fmol/mg protein, n = 8). In competition studies, ICI-89406 (beta 1-antagonist) and ICI-118551 (beta 2-antagonist) bound with high affinity, fitting a two-site model. Isoproterenol increased intracellular adenosine 3',5'-cyclic monophosphate (cAMP) accumulation (half-maximal effective concentration = 200 nM). Propranolol completely inhibited isoproterenol-induced cAMP accumulation [half-maximal inhibitory concentration (IC50) = 270 nM]. ICI-89406 and ICI-118551 inhibited cAMP accumulation by 50% (IC50 = 1.5 microM and 1.7 microM, respectively). The combined addition of ICI-89406 and ICI-118551 resulted in a curve indistinguishable from that of propranolol. The beta 1- and beta 2-adrenoceptor mRNAs have been demonstrated using reverse transcription-polymerase chain reaction. In initial and terminal IMCD cells, propranolol (3 microM) inhibited isoproterenol-stimulated cAMP accumulation by 80%, whereas ICI-89406 (3 microM) and ICI-118551 (3 microM) resulted in only partial inhibition (50%). We conclude that both beta 1- and beta 2-adrenoceptors are expressed in initial and terminal IMCD cells in primary culture.


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D. P. Wallace, G. Reif, A.-M. Hedge, J. B. Thrasher, and P. Pietrow
Adrenergic regulation of salt and fluid secretion in human medullary collecting duct cells
Am J Physiol Renal Physiol, October 1, 2004; 287(4): F639 - F648.
[Abstract] [Full Text] [PDF]


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