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Am J Physiol Renal Physiol 281: F172-F178, 2001;
0363-6127/01 $5.00
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Vol. 281, Issue 1, F172-F178, July 2001

alpha 1-Adrenoceptor subtypes on rat afferent arterioles assessed by radioligand binding and RT-PCR

Max Salomonsson, Melinda Oker, Susan Kim, Hua Zhang, James E. Faber, and William J. Arendshorst

Department of Cell and Molecular Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599-7545

We utilized [3H]prazosin saturation and competition radioligand binding studies to characterize the expression of alpha 1-adrenoceptors in preglomerular vessels. mRNA for adrenoceptor subtypes was assayed using RT-PCR. The vessels were isolated using an iron oxide-sieving method. [3H]prazosin bound to a single class of binding sites (Kd 0.087 ± 0.012 nM, Bmax 326 ± 56 fmol/mg protein). Phentolamine displaced [3H]prazosin (0.2 nM) with a pKi of 8.37 ± 0.09. Competition with the selective alpha 1A-adrenoceptor antagonist 5-methylurapidil fit a two-site model (pKi 9.38 ± 0.21 and 7.04 ± 0.15); 59 ± 3% of the sites were high-affinity, and 41 ± 3% were low-affinity binding sites. Competition with the alpha 1D-adrenoceptor antagonist 8-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-8-azaspiro[4.5]decane-7,9-dione dihydrochloride (BMY-7378) fit a one-site model with low affinity (pKi 6.83 ± 0.03). The relative contents of alpha 1A-, alpha 1B-, and alpha 1D-adrenoceptor mRNAs were 64 ± 5, 25 ± 5, and 11 ± 1%, respectively. Thus there was a very good correlation between mRNA and receptor binding for the subtypes. These data indicate a predominance of the alpha 1A-adrenoceptor subtype in rat renal resistance vessels, with smaller densities of alpha 1B- and alpha 1D-adrenoceptors.

renal circulation; vascular smooth muscle; reverse transcription-polymerase chain reaction; mRNA; renal nerve; catecholamine


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