title: Novel splice variants of rat CaV2.1 that lack much of the synaptic protein interaction site are expressed in neuroendocrine cells.
abstract: Voltage-gated Ca(2+) channels are responsible for the activation of the Ca(2+) influx that triggers exocytotic secretion. The synaptic protein interaction (synprint) site found in the II-III loop of Ca(V)2.1 and Ca(V)2.2 mediates a physical association with synaptic proteins that may be crucial for fast neurotransmission and axonal targeting. We report here the use of nested PCR to identify two novel splice variants of rat Ca(V)2.1 that lack much of the synprint site. Furthermore, we compare immunofluorescence data derived from antibodies directed against sequences in the Ca(V)2.1 synprint site and carboxyl terminus to show that channel variants lacking a portion of the synprint site are expressed in two types of neuroendocrine cells. Immunofluorescence data also suggest that such variants are properly targeted to neuroendocrine terminals. When expressed in a mammalian cell line, both splice variants yielded Ca(2+) currents, but the variant containing the larger of the two deletions displayed a reduced current density and a marked shift in the voltage dependence of inactivation. These results have important implications for Ca(V)2.1 function and for the mechanisms of Ca(V)2.1 targeting in neurons and neuroendocrine cells.
authors.affiliation: Department of Physiology, College of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5.
authors.forename: W R A Kosala J S
authors.initials: WR
authors.lastname: Rajapaksha
authors.forename: Daoyi
authors.initials: D
authors.lastname: Wang
authors.forename: Jonathan N
authors.initials: JN
authors.lastname: Davies
authors.forename: Lina
authors.initials: L
authors.lastname: Chen
authors.forename: Gerald W
authors.initials: GW
authors.lastname: Zamponi
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Fisher
mesh_terms: D017398:Alternative Splicing / Q000502:physiology*; D000818:Animals; D001369:Axons / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D020864:Calcium Channels, N-Type / Q000096:biosynthesis* / Q000235:genetics; D023902:Chromosome Pairing / Q000502:physiology*; D008297:Male; D009490:Neurosecretory Systems / Q000166:cytology / Q000378:metabolism*; D016716:PC12 Cells; D017433:Protein Structure, Secondary / Q000502:physiology; D051381:Rats; D020318:Rats, Long-Evans
chemical_list: D002118:Calcium; D020864:Calcium Channels, N-Type; C503279:Cacna1b protein, rat