Quantification of two splicing events in the L-type calcium channel alpha-1 subunit of intestinal smooth muscle and other tissues

cDNA fragments encoding a representative region of the L-type calcium channel alpha-1 subunit of rabbit intestine smooth muscle were amplified by polymerase chain reaction (PCR). The nucleotide sequences of these intestine clones shared a high similarity with aorta, lung and heart calcium channels. However, in the extracellular loop between the third and fourth segments of domain IV and in the transmembrane IVS3 segment itself, we observed primary sequence variations corresponding to alternative splicing phenomenons. Since structural differences of L-type calcium channel alpha-1 subunits could result in functional variations, the respective expression frequency of these isoforms was determined in various tissues and species, and in the embryonic A7r5 cell line. The ontogeny of these splicing events was also examined from tissues of different ages. From this quantitative study, carried out by PCR of reverse-transcribed mRNA, it clearly appears that the observed splicing processes in the IVS3-IVS4 region are not only tissue-dependent but also regulated during development