IS THE M₁‐MUSCARINIC RECEPTOR A CANDIDATE GENE FOR HYPERTENSION IN THE SPONTANEOUSLY HYPERTENSIVE RAT?

Abstract

1. Central nervous system muscarinic receptors have been implicated in genetic models of hypertension as well as in stress adaptive cardiovascular responses. To determine if the sequence of the MI-muscarinic receptor is pathogenetic for hypertension, we analysed the differences between SHRLJ (spontaneously hypertensive rats) and WKYLJ (Wistar-Kyoto) rats in the sequence of the MI-receptor gene.2. Specific primer sets were synthesized so as to cover the gene in 10 fragments (158-344 base pairs) with overlaps. Analysis was conducted by both polymerase chain reaction (PCR)-acrylamide and by single-stranded conformation polymorphism (SSCP).3. No polymorphic locus was found by non-denaturing PCR-acrylamide nor by denaturing SSCP. Slight intrastrain variations in fragment 2 of the parental strains were examined by direct sequencing; however, no difference in sequence was found between SHRLJ and WKYLI. To identify the chromosomal location and possible linkage with a polymorphic locus, rat/mouse somatic hybrid cell lines were examined. The rat MI-muscarinic receptor was assigned to chromosome 1. 4.Based on analysis of the coding sequence of the gene, our data do not support the hypothesis that the MI-muscarinic receptor is a candidate gene for hypertension. We conclude that, pending analyses of the promoter and regulatory regions of the gene, differences in the MI-muscarinic receptor in SHR either in receptor expression or in physiological response must be due to altered intracellular signalling or extracellular transynaptic events, but not due to a mutation of the gene sequence.