IDENTIFICATION OF THE ALPHA(1C)-ADRENOCEPTOR IN RABBIT ARTERIES AND THE HUMAN SAPHENOUS-VEIN USING THE POLYMERASE CHAIN-REACTION

The expression of the alpha(1C)-adrenoceptor subtype in human and rabbit blood vessels has been analyzed using the reverse transcriptase/polymerase chain reaction technique (RT/PCR). The 20 bp primers employed were designed from the bovine alpha(1C)-adrenoceptor and flank a least conserved region - the putative third cytoplasmic loop. RT/PCR products generated from rabbit and human brain mRNA both had 93% homology to the bovine alpha(1C)-adrenoceptor and were used as species and subtype specific probes in Southern blot analysis of vascular RT/PCR products. Poly A(+) RNA was purified from the human saphenous vein and rabbit aorta, renal, pulmonary and central ear arteries and amplified by RT/PCR. Size analysis by agarose gel electrophoresis, together with Southern hybridization of the resulting cDNA products confirm the expression of the alpha(1C)-adrenoceptor in these vessels.