title: Expression of Calcium Channel Subunit Variants in Small Mesenteric Arteries of WKY and SHR.
abstract: BACKGROUND
  Enhanced function of dihydropyridine-sensitive Ca2+ channels (CaV) in hypertensive arterial myocytes (HAM) is well accepted. Increased protein expression of pore forming α1-subunits contributes to this effect, but cannot explain all of the differences in CaV properties in HAM. We hypothesized that differences in expression of CaV subunits and/or their splice variants also contribute.
  
  
  METHODS
  RNA, protein, and myocytes were isolated from small mesenteric arteries (SMA) of 20-week-old male WKY and SHR and analyzed by polymerase chain reaction (PCR), sequencing, immunoblotting, and patch clamp methods.
  
  
  RESULTS
  Cav1.2 α1, β2c, and α2δ1d were the dominant subunits expressed in both WKY and SHR with a smaller amount of β3a. Real-time PCR indicated that the mRNA abundance of β3a and α2δ1 but not total Cav1.2 α1 or β2c were significantly larger in SHR. Analysis of alternative splicing of Cav1.2 α1 showed no differences in abundance of mutually exclusive exons1b, 8, 21 and 32 or alternative exons33 and 45. However, inclusion of exon9* was higher and a 73 nucleotide (nt) deletion in exon15 (exon15Δ73) was lower in SHR. Immunoblot analysis showed higher protein levels of Cav1.2 α1 (1.61±0.05), β3 (1.80±0.32), and α2δ1 (1.80±0.24) but not β2 in SHR.
  
  
  CONCLUSIONS
  The lower abundance of exon15Δ73 transcripts in SHR results in a larger fraction of total Cav1.2 mRNA coding for full-length CaV protein, and the higher abundance of exon9* transcripts and CaVβ3a protein likely contribute to differences in gating and kinetics of CaV currents in SHR. Functional studies of Ca2+ currents in native SMA myocytes and HEK cells transiently transfected with CaV subunits support these conclusions.
authors.affiliation: Program in Cardiovascular Disease, Lankenau Institute for Medical Research, Wynnewood, Pennsylvania. coxr@mlhs.org.
authors.forename: Robert H
authors.initials: RH
authors.lastname: Cox
authors.affiliation: Program in Cardiovascular Disease, Lankenau Institute for Medical Research, Wynnewood, Pennsylvania.
authors.forename: Samantha
authors.initials: S
authors.lastname: Fromme
mesh_terms: D000818:Animals; D015220:Calcium Channels / Q000378:metabolism*; D020869:Gene Expression Profiling; D006973:Hypertension / Q000378:metabolism*; D008297:Male; D008638:Mesenteric Arteries / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D020033:Protein Isoforms / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY
chemical_list: D015220:Calcium Channels; D020033:Protein Isoforms; D021122:Protein Subunits
keywords: Ca2+; alternative splicing; blood pressure; channel subunits; channels.; gene expression; hypertension; hypertensive rats; protein abundance; small mesenteric arteries; voltage gated Ca2+
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