title: Alternative splicing generates a smaller assortment of CaV2.1 transcripts in cerebellar Purkinje cells than in the cerebellum.
abstract: P/Q-type calcium channels control many calcium-driven functions in the brain. The CACNA1A gene encoding the pore-forming CaV2.1 (alpha1A) subunit of P/Q-type channels undergoes alternative splicing at multiple loci. This results in channel variants with different phenotypes. However, the combinatorial patterns of alternative splice events at two or more loci, and hence the diversity of CaV2.1 transcripts, are incompletely defined for specific brain regions and types of brain neurons. Using RT-PCR and splice variant-specific primers, we have identified multiple CaV2.1 transcript variants defined by different pairs of splice events in the cerebellum of adult rat. We have uncovered new splice variations between exons 28 and 34 (some of which predict a premature stop codon) and a new variation in exon 47 (which predicts a novel extended COOH-terminus). Single cell RT-PCR reveals that each individual cerebellar Purkinje neuron also expresses multiple alternative CaV2.1 transcripts, but the assortment is smaller than in the cerebellum. Two of these variants encode different extended COOH-termini which are not the same as those previously reported in Purkinje cells of the mouse. Our patch-clamp recordings show that calcium channel currents in the soma and dendrites of Purkinje cells are largely inhibited by a concentration of omega-agatoxin IVA selective for P-type over Q-type channels, suggesting that the different transcripts may form phenotypic variants of P-type calcium channels in Purkinje cells. These results expand the known diversity of CaV2.1 transcripts in cerebellar Purkinje cells, and propose the selective expression of distinct assortments of CaV2.1 transcripts in different brain neurons and species.
authors.affiliation: Department of Pharmacology, University of Bristol, Bristol, United Kingdom.
authors.forename: Srinivasan
authors.initials: S
authors.lastname: Kanumilli
authors.forename: Elizabeth W
authors.initials: EW
authors.lastname: Tringham
authors.forename: C Elizabeth
authors.initials: CE
authors.lastname: Payne
authors.forename: Jonathan R B
authors.initials: JR
authors.lastname: Dupere
authors.forename: Kanamarlapudi
authors.initials: K
authors.lastname: Venkateswarlu
authors.forename: Maria M
authors.initials: MM
authors.lastname: Usowicz
mesh_terms: D017398:Alternative Splicing / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D015220:Calcium Channels / Q000187:drug effects / Q000235:genetics; D020864:Calcium Channels, N-Type / Q000737:chemistry / Q000235:genetics*; D002531:Cerebellum / Q000166:cytology* / Q000378:metabolism*; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D005091:Exons / Q000235:genetics; D005838:Genotype; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011689:Purkinje Cells / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020867:omega-Agatoxin IVA / Q000494:pharmacology
chemical_list: D015220:Calcium Channels; D020864:Calcium Channels, N-Type; D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D020867:omega-Agatoxin IVA; C491367:Cacna1a protein, rat