Cloning of rabbit alpha(1b)-adrenoceptor and pharmacological comparison of alpha(1a)-, alpha(1b)- and alpha(1d)-adrenoceptors in rabbit.

We have isolated a cDNA clone of the rabbit alpha(1b)-adrenoceptor which has an open reading frame of 1557 nucleotides encoding a protein of 518 amino acids. The sequence shows higher identity to those of hamster, human, and rat alpha(1b)-adrenoceptors than to those of rabbit alpha(1a)- and alpha(1d)-adrenoceptors. The pharmacological binding properties of this clone expressed in Cos-7 cells showed a characteristic profile as alpha(1b)-adrenoceptor; high affinity for prazosin (pK(i)=10.3), relatively high affinity for tamsulosin (9.5) and low affinity for (-)-(R)-1-(3-hydroxypropyl)-5-[2-[[2-[2-(2,2, 2-trifluoroethoxy)phenoxy]ethyl]amino]propyl]indoline-7-carboxamid e (KMD3213) (8.5), 2-(2,6-dimethoxy-phenoxyethyl)-aminomethyl-1, 4-benzodioxane hydrochloride (WB4101) (8.7), and 8-[2-[4-(2-methoxy-phenyl)-L-piperazinyl]-8-azaspiro[4,5]decane-7, 9-dione dihydrochloride (BMY7378) (7.3). We have compared the levels of mRNA expression of three alpha(1)-adrenoceptor subtypes in rabbit tissues using the competitive reverse transcription/polymerase chain reaction (RT/PCR) assay. In most rabbit tissues except heart, alpha(1a)-adrenoceptor mRNA was expressed 10 folds more than the other two subtypes. However, binding experiments with [3H]prazosin and [3H]KMD3213 in rabbit tissues revealed a poor relationship between binding density and mRNA level. Especially, alpha(1b) binding sites were exclusively predominant in spleen, whereas the alpha(1b) subtype was minor at the mRNA level. These results indicate a high identity of structural and pharmacological profiles of three distinct alpha(1)-adrenoceptor subtypes between rabbit and other species, but there are species differences in their distribution.