Skip to main content
Log in

Developmental regulation of the L-type calcium channel α1C subunit expression in heart

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

We used Northern analyses, RNase protection assays and immunoblot analyses to examine the relationship among developmental age of the heart, abundance of mRNA and L-type calcium channel α1Csubunit protein, and to establish the size of the native protein in heart. Northern analysis, RNase protection assays, and immunoblots were used to study RNA and protein from rat heart of various ages. In fetal and adult ventricles there was a predominant 8.3-kb transcript for the α1C subunit with no change in transcript size during development. RNase protection assays demonstrated a 2-fold increase in abundance of the DHP receptor message during postnatal development. Immunoblots identified a 240 kD protein, corresponding to the predicted molecular mass of the full length α1C subunit. No change in size of protein for the α1C subunit was observed at any developmental stage and there was no evidence for a truncated isoform. There was an approximate 2-fold increase in α1C subunit protein in ventricular homogenates during postnatal development. Thus, in the developing rat heart, alterations in calcium channel properties during development appear to result neither from alternative splicing that produces a smaller transcript for the α1C subunit nor from expression of a truncated protein, but at least in part from transcriptionally-regulated expression of the 240 kDa polypeptide.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Marsh JD, Allen PD: Developmental regulation of cardiac calcium channels and contractile sensitivity to [Ca]o. Am J Physiol 256: H179–H185, 1989

    PubMed  Google Scholar 

  2. Tohse N, Masuda H, Sperelakis N: Novel isoform of Ca2+ channel in rat fetal cardiomyocytes. J Physiol (London) 451: 295–306, 1992

    PubMed  Google Scholar 

  3. Huynh TV, Chen F, Wetzel GT, Friedman WF, Klitzner TS: Developmental changes in membrane calcium and potassium currents in fetal, neonatal and adult rabbit ventricular myocytes. Circ Res 70: 508–15, 1992

    PubMed  Google Scholar 

  4. Artman M: Sarcolemmal Na-Ca exchange activity and immunoreactivity in developing rabbit hearts. Am J Physiol 263: H1506–H1513, 1992

    PubMed  Google Scholar 

  5. Brillantes A-MB, Bezprozvannaya S, Marks AR: Developmental and tissue-specific regulation of rabbit skeletal and cardiac muscle calcium channels involved in excitation-contraction coupling. Circ Res 75: 503–10, 1994

    PubMed  Google Scholar 

  6. Lompre AM, Lambert F, Lakatta EG, Schwartz K: Expression of sarcoplasmic reticulum Ca(2+)-ATPase and calsequestrin genes in rat heart during ontogenic development and aging. Circ Res 69: 1380–388, 1991

    PubMed  Google Scholar 

  7. Ogawa S, Barnett JV, Sen L, Galper JB, Smith TW, Marsh JD: Direct contact between sympathetic neurons and rat cardiac myocytes in vitro increases expression of functional calcium channels. J Clin Invest 89: 1085–93, 1992

    PubMed  Google Scholar 

  8. Kim D, Marsh JD, Smith TW: Effects of thyroid hormone on slow Ca channel function in cultured chick ventricular cells. J Clin Invest 80: 88–94, 1987

    PubMed  Google Scholar 

  9. Marsh JD: Co-regulation of calcium channels and beta-adrenergic receptors in cultured chick embryo ventricular cells. J Clin Invest 84: 817–23, 1989

    PubMed  Google Scholar 

  10. Haase H, Kresse A, Hohaus A, Schulte HD, Maier M, Osterziel KJ, Lange PE, Morano I: Expression of calcium channel subunits in the normal and diseased human myocardium. J Mol Med 74: 99–104, 1996

    PubMed  Google Scholar 

  11. Cohen NM, Lederer WJ: Changes in the calcium current of rat heart ventricular myocytes during development. J Physiol (London) 406: 115–146, 1988

    Google Scholar 

  12. Aiba S, Creazzo TL: Comparison of the number of dihydropyridine receptors with the number of functional L-type calcium channels in embryonic heart. Circ Res 72: 396–402, 1993

    PubMed  Google Scholar 

  13. Diebold RJ, Koch WK, Ellinor PT, Wang JJ, Muthuchamy M, Wieczorek DF, Schwartz A: Mutually exclusive exon splicing of the cardiac calcium channel alpha1 subunit gene generates developmentally regulated isoforms in the rat heart. Proc Natl Acad Sci USA 89: 1497–501, 1992

    PubMed  Google Scholar 

  14. Hosey MM, Chang FC, O'Callahan CM, Ptasienski J. L-type calcium channels in cardiac and skeletal muscle. Purification and phosphorylation. Ann N Y Acad Sci 560: 27–38, 1989

    PubMed  Google Scholar 

  15. Mitterdorfer J, Froschmayr M, Grabner M, Moebius FF, Glossman H, Striessnig J: Identification of PK-A phosphorylation sites in the carboxyl terminus of L-type calcium channel alpha1 subunits. Biochem 35: 9400–9406, 1996

    Article  Google Scholar 

  16. De Jongh KS, Murphy BJ, Colvin AA, Hell JW, Takahashi M, Catterall WA: Specific phosphorylation of a site in the full-length form of the alpha 1 subunit of the L-type calcium channel by adenosine 3′5′-cyclic monophosphate-dependent protein kinase. Biochem 35: 10392–10402, 1996

    Article  Google Scholar 

  17. Yoshida A, Takahashi M, Nishimura S, Takeshima H, Kokubun S: Cyclic AMP-dependent phosphorylation and regulation of the cardiac dihydropyridine-sensitive calcium channel. FEBS Lett 309: 343–349, 1992

    Article  PubMed  Google Scholar 

  18. Schmidt JW, Catterall WA: Biosynthesis and processing of the alpha subunit of the voltage sensitive sodium channel in rat brain neurons. Cell 46: 437–445, 1986

    Article  PubMed  Google Scholar 

  19. De Jongh KS, Warner C, Colvin AA, Catterall WA. Characterization of the two size forms of the alpha 1 subunit of skeletal muscle L-type calcium channels. Proc Natl Acad Sci. USA 88: 10778–10782, 1991

    PubMed  Google Scholar 

  20. Hell JW, Yokoyama CT, Wong ST, Warner C, Snutch TP, Catterall WA: Differential phosphorylation of two size forms of the neuronal class C L-type calcium channel alpha1 subunit. J Biol Chem 268: 1945–57, 1993

    Google Scholar 

  21. Liu L, Poornima I, H'Hara DS, Hines RN, Marsh JD: DHP receptor mRNA levels and protein abundance are developmentally regulated in rat ventricle. Circulation 92: I372, 1995

    Google Scholar 

  22. Farrell, RE: RNA Methodologies: A Laboratory Guide for Isolation and Characterization. Academic Press; New York 1993

    Google Scholar 

  23. Church GM, Gilbert W: Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995, 1984

    PubMed  Google Scholar 

  24. Mikami A, Imoto K, Tanabe T, Niidome T, Mori Y, Takeshima H, Narumiya S, Numa S: Primary structure and functional expression of the cardiac dihydropyridine-sensitive calcium channel. Nature 340: 230–233, 1989

    Article  PubMed  Google Scholar 

  25. Sanger F, Nicklen S, Coulson AR: DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467, 1977

    PubMed  Google Scholar 

  26. Yaney GC, Wheeler MB, Wei XY, Perez-Reyes E, Birnbaumer L, Boyd AB, Moss LG: Cloning of a novel alpha-1 subunit of the voltage-dependent calcium channel from the beta cell. Mol Endocrinol 6: 2143–2152, 1992

    Article  PubMed  Google Scholar 

  27. Wyatt CN, Campbell V, Brodbeck J, Brice NL, Page KN, Berrow NS, Brickley K, Terracciano CM, Naqvi RU, MacLeod KT, et al.: Voltagedependent binding and calcium channel current inhibition by an antialpha1-D subunit antibody in rat dorsal root ganglion neurons and guinea-pig myocytes. J Physiol 502: 307–319, 1997

    Article  PubMed  Google Scholar 

  28. Cala SE, Miles K: Phosphorylation of the cardiac isoforms of calsequestrin in cultured rat myotubes and in rat skeletal muscle. Biochim Biophys Acta 1118: 277–279, 1992

    PubMed  Google Scholar 

  29. Davidoff AJ, Maki TM, Ellingsen O, Marsh JD: Expression of calcium channels in adult cardiac myocytes is regulated by calcium. J Mol Cell Cardiol 29: 1791–1803, 1997

    Article  PubMed  Google Scholar 

  30. Takahashi T, Allen PD, Lacro RV, Marks AR, Dennis AR, Schoen FJ, Grossman W, Marsh JD, Izumo S: Expression of dihydropyridine receptor (Ca2+ channel) and calsequestrin genes in the myocardium of patients with end-stage heart failure. J Clin Invest 90: 927–935, 1992

    PubMed  Google Scholar 

  31. Wei X, Neely A, Lacerda AE, Olcese R, Stefani E, Perez-Reyes E, Birnbaumer L: Modification of Ca channel activity by deletions at the carboxyl terminus of the cardiac alpha-1 subunit. J Biol Chem 269: 1635–1640, 1994

    PubMed  Google Scholar 

  32. Soldatov NM: Genomic structure of the L-type calcium channel. Genomics 22: 77–87, 1994

    Article  PubMed  Google Scholar 

  33. Soldatov NM, Bouron A, Reuter H: Different voltage-dependent inhibition by dihydropyridines of human Ca channel splice variants. J Biol Chem 270: 10540–10543, 1995

    Article  PubMed  Google Scholar 

  34. Wetzel GT, Chen F, Klitzner TS: L-and T-type calcium channels in acutely isolated neonatal and adult cardiac myocytes. Pediatr Res 30: 89–94, 1991

    PubMed  Google Scholar 

  35. Chien AJ, Zhao X, Shirokov RE, Puri TS, Chang CF, Sun D, Rios E, Hosey MM: Roles of a membrane-localized beta subunit in the formation and targeting of functional L-type Ca2+ channels. J Biol Chem 270: 30036–30044, 1995

    PubMed  Google Scholar 

  36. Chang FC, Hosey MM: Dihydropyridine and phenylalkylamine receptors associated with cardiac and skeletal muscle calcium channels are structurally different. J Biol Chem 263: 929–937, 1988

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leu, L., O'Hara, D.S., Cala, S.E. et al. Developmental regulation of the L-type calcium channel α1C subunit expression in heart. Mol Cell Biochem 205, 101–109 (2000). https://doi.org/10.1023/A:1007013900827

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1007013900827

Navigation