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Regulation of Muscle Cav1.1 Channels by Long-term Depolarization Involves Proteolysis of the α1s Subunit

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Abstract

The effects of long-term depolarization on frog skeletal muscle Cav1.1 channels were assessed. Voltage-clamp and Western-blot experiments revealed that long-term depolarization brings about a drastic reduction in the amplitude of currents flowing through Cav1.1 channels and in the levels of the α1s subunit, the main subunit of muscle L-type channels. The decline of both phenomena was prevented by the action of the protease inhibitors E64 (50 μM) and leupeptin (50 μM). In contrast, long-term depolarization had no effect on β1, the auxiliary subunit of α1s. The levels of mRNAs coding the α1s and the β1 subunits were measured by RNase protection assays. Neither the content of the α1s nor the β1 subunit mRNAs were affected by long-term depolarization, indicating that the synthesis of Cav1.1 channels remained unaffected. Taken together, our experiments suggest that the reduction in the amplitude of membrane currents and in the α1s subunit levels is caused by increased degradation of this subunit by a Ca2+-dependent protease.

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References

  • J. Arreola J. Calvo M.C. Garcia J.A. Sanchez (1987) ArticleTitleModulation of calcium channels of twitch skeletal muscle fibres of the frog by adrenaline and cyclic adenosine monophosphate J. Physiol 393 307–330 Occurrence Handle1:CAS:528:DyaL1cXhs1Kmuw%3D%3D Occurrence Handle2451739

    CAS  PubMed  Google Scholar 

  • D.E. Atsma E.M. Bastiaanse A. Jerzewski L.J. Valk ParticleVan der A. Laarse ParticleVan der (1995) ArticleTitleRole of calcium-activated neutral protease (Calpain) in cell death in cultured neonatal rat cardiomyocytes during metabolic inhibition Circ. Res 76 1071–1078 Occurrence Handle1:CAS:528:DyaK2MXlvVSht7w%3D Occurrence Handle7758161

    CAS  PubMed  Google Scholar 

  • B. Belles J. Hescheler W. Trautwein K. Blomgren J.O. Karlsson (1988) ArticleTitleA possible physiological role of the Ca-dependent protease calpain and its inhibitor calpastatin on the Ca current in guinea pig myocytes Pfluegers Arch 412 554–556 Occurrence Handle10.1007/BF00582548 Occurrence Handle1:CAS:528:DyaL1cXlvVOrsbs%3D

    Article  CAS  Google Scholar 

  • D.M. Bers (2002) ArticleTitleCardiac excitation-contraction coupling Nature 415 198–205 Occurrence Handle10.1038/415198a Occurrence Handle1:CAS:528:DC%2BD38Xms1Whsg%3D%3D Occurrence Handle11805843

    Article  CAS  PubMed  Google Scholar 

  • Carrillo E., Galindo J.M., García M.C., Sánchez J.A. 2003. Regulación de los canales de calcio Cav1.1: Degradación por proteasas activadas por el influjo de Ca2+ a través del canal. In: Memorias XLVl Congreso Nacional de Ciencias Fisiológicas, O19, Aguascalientes, México

  • W.A. Catterall K. Jongh ParticleDe E. Rotman J. Hell R. Westenbrock S.J. Dubel T.P. Snutch (1993) ArticleTitleMolecular properties of calcium channels in skeletal muscle and neurons Ann. N.Y. Acad. USA 681 342–355 Occurrence Handle1:CAS:528:DyaK3sXltFCjtro%3D

    CAS  Google Scholar 

  • J.E. Chad R. Eckert (1986) ArticleTitleAn enzymatic mechanism for calcium current inactivation in dialysed Helix neurones J. Physiol 378 31–51 Occurrence Handle1:CAS:528:DyaL28XkslKlu7w%3D Occurrence Handle2432251

    CAS  PubMed  Google Scholar 

  • P. Chomczynski N. Sacchi (1987) ArticleTitleSingle-step method of RNA isolation by acid-guanidinium thiocyanate-phenol-chloroform extraction Anal. Biochem 162 156–159 Occurrence Handle1:CAS:528:DyaL2sXitFSns7Y%3D Occurrence Handle2440339

    CAS  PubMed  Google Scholar 

  • D.E. Croall G.N. DeMartino (1991) ArticleTitleCalcium-activated neutral protease (calpain) system: structure, function, and regulation Physiol. Rev 71 813–847 Occurrence Handle1:CAS:528:DyaK3MXltFWjur4%3D Occurrence Handle2057527

    CAS  PubMed  Google Scholar 

  • W.R. Dayton J.V. Schollmeyer (1981) ArticleTitleImmunocytochemical localization of a calcium-activated protease in skeletal muscle cells Exp. Cell. Res 136 423–433 Occurrence Handle10.1016/0014-4827(81)90022-7 Occurrence Handle1:CAS:528:DyaL38XhtF2lt7o%3D Occurrence Handle6273198

    Article  CAS  PubMed  Google Scholar 

  • K.S. Jongh ParticleDe D.K. Merrick W.A. Catterall (1989) ArticleTitleSubunits of purified calcium channels: a 212-kDa form of alpha 1 and partial amino acid sequence of a phosphorylation site of an independent beta subunit Proc. Natl. Acad. Sci. USA 86 8585–8589 Occurrence Handle2554320

    PubMed  Google Scholar 

  • E.M. Lorme ParticleDe C.S. Rabe R. McGee (1988) ArticleTitleRegulation of the number of functional voltage-sensitive Ca2+ channels on PC12 cells by chronic changes in membrane potential J. Pharmacol 244 838–843

    Google Scholar 

  • E.A. Ertel K.P. Campbell M.M. Harpold F. Hofmann Y. Mori E. Perez-Reyes A. Schwartz T.P. Snutch T. Tanabe L. Birnbaumer R.W. Tsien W.A. Catterall (2000) ArticleTitleNomenclature of voltage-gated calcium channels Neuron 25 533–535 Occurrence Handle10.1016/S0896-6273(00)81057-0 Occurrence Handle1:CAS:528:DC%2BD3cXisFKrtL0%3D Occurrence Handle10774722

    Article  CAS  PubMed  Google Scholar 

  • J. Escamilla J.M. Farias R. Garcia M.C. Garcia J.A. Sánchez (2001) ArticleTitleLong-term depolarization regulates the α1s subunit of skeletal muscle Ca2+ channels and the amplitude of L-type Ca2+ currents Pfluegers Arch 442 882–890 Occurrence Handle10.1007/s004240100606 Occurrence Handle1:CAS:528:DC%2BD3MXmslOktL0%3D

    Article  CAS  Google Scholar 

  • O. Feron T. Godfraind (1995) ArticleTitleRegulation of the L-type calcium channel alpha-1 subunit by chronic depolarization in the neuron-like PC12 and aortic smooth muscle A7r5 cell lines Pfluegers Arch 430 323–332 Occurrence Handle10.1007/BF00373906 Occurrence Handle1:CAS:528:DyaK2MXmvFamsLY%3D

    Article  CAS  Google Scholar 

  • M. Fosset E. Jaimovich E. Delpont M. Lazdunski (1983) ArticleTitle[3H]nitrendipine receptors in skeletal muscle J. Biol. Chem 258 6086–6092 Occurrence Handle1:CAS:528:DyaL3sXktFKgsbY%3D Occurrence Handle6304026

    CAS  PubMed  Google Scholar 

  • J.L. Franklin D.J. Fickbohm A.L. Willard (1992) ArticleTitleLong-term regulation of normal calcium currents by prolonged changes of membrane potential J. Neurosci 12 1726–1735 Occurrence Handle1:CAS:528:DyaK38XktVCqu7o%3D Occurrence Handle1315850

    CAS  PubMed  Google Scholar 

  • V. Ganitkevich V. Hasse G. Pfitzer (2002) ArticleTitleCa2+-dependent and Ca2+-independent regulation of smooth muscle contraction J. Muscle Res. Cell. Motil 23 47–52 Occurrence Handle10.1023/A:1019956529549 Occurrence Handle1:CAS:528:DC%2BD38Xms1ektL4%3D Occurrence Handle12363284

    Article  CAS  PubMed  Google Scholar 

  • D.E. Goll V.F. Thompson H.Q. Li W. Wei J. Cong (2003) ArticleTitleThe calpain system Physiol. Rev 83 731–801 Occurrence Handle1:CAS:528:DC%2BD3sXmtVGmsL8%3D Occurrence Handle12843408

    CAS  PubMed  Google Scholar 

  • R.G. Gregg A. Messing C. Strube M. Beurg R. Moss M. Behan M. Sukhareva S. Haynes J.A. Powell R. Coronado P.A. Powers (1996) ArticleTitleAbsence of the β subunit (cchb1) of the skeletal muscle dihydropyridine receptor alters expression of the α1 subunit and eliminates excitation-contraction coupling Proc. Natl. Acad. Sci. USA 93 13961–13966 Occurrence Handle10.1073/pnas.93.24.13961 Occurrence Handle1:CAS:528:DyaK28Xnt1Gltb8%3D Occurrence Handle8943043

    Article  CAS  PubMed  Google Scholar 

  • L.Y. Hao A. Kameyama S. Kuroki J. Takano E. Takano M. Maki M. Kameyama (2000) ArticleTitleCalpastatin domain L is involved in the regulation of L-type Ca2+ channels in guinea pig cardiac myocytes Biochem. Biophys. Res. Commun 279 756–761 Occurrence Handle10.1006/bbrc.2000.4040 Occurrence Handle1:CAS:528:DC%2BD3MXivVaktg%3D%3D Occurrence Handle11162425

    Article  CAS  PubMed  Google Scholar 

  • B. Hille D.T. Campbell (1976) ArticleTitleAn improved Vaseline gap voltage clamp for skeletal muscle fibers J. Gen. Physiol 67 265–293 Occurrence Handle10.1085/jgp.67.3.265 Occurrence Handle1:STN:280:CSmC2s%2FptlU%3D Occurrence Handle1083424

    Article  CAS  PubMed  Google Scholar 

  • B. Hille (2001) Ion Channels of Excitable Membranes Sinauer Associates, Inc USA

    Google Scholar 

  • K. Kinbara H. Sorimachi S. Ishiura K. Suzuki (1998) ArticleTitleSkeletal muscle-specific calpain, p94 Biochem. Pharmacol 56 415–420 Occurrence Handle10.1016/S0006-2952(98)00095-1 Occurrence Handle1:CAS:528:DyaK1cXlsVCrurg%3D Occurrence Handle9763216

    Article  CAS  PubMed  Google Scholar 

  • T. Kumamoto W.C. Kleese J. Cong D.E. Goll P.R. Pierce R.E. Allen (1992) ArticleTitleLocalization of the Ca2+-dependent proteinases and their inhibitor in normal, fasted and denervated rat skeletal muscle Anat. Rec 232 60–77 Occurrence Handle10.1002/ar.1092320108 Occurrence Handle1:CAS:528:DyaK38XhsVyrsLo%3D Occurrence Handle1536466

    Article  CAS  PubMed  Google Scholar 

  • G.D. Lamb (1992) ArticleTitleDHP receptors and excitation-contraction coupling J. Muscle Res. Cell Motil 13 394–405 Occurrence Handle10.1007/BF01738035 Occurrence Handle1:CAS:528:DyaK38XmtlSlt7s%3D Occurrence Handle1328292

    Article  CAS  PubMed  Google Scholar 

  • J. Liu R. Bangalore A. Rutledge D.J. Triggle (1994) ArticleTitleModulation of L-type Ca2+ channels in clonal rat pituitary cells by membrane depolarization Mol. Pharmacol 45 1198–1206 Occurrence Handle1:CAS:528:DyaK2cXksl2ntL4%3D Occurrence Handle8022413

    CAS  PubMed  Google Scholar 

  • A. Liu J. Rutledge D.J. Triggle (1995) ArticleTitleShort-term regulation of neuronal calcium channels by depolarization Ann. N.Y. Acad. Sci. USA 765 119–133 Occurrence Handle1:CAS:528:DyaK28XltFajsw%3D%3D

    CAS  Google Scholar 

  • W. Melzer A. Hermann-Frank H.C.H. Lüttgau (1995) ArticleTitleThe role of Ca2+ ions in excitation-contraction coupling of skeletal muscle fibres Biochim. Biophys. Acta 1241 59–116 Occurrence Handle1:CAS:528:DyaK2MXmsVahs7w%3D Occurrence Handle7742348

    CAS  PubMed  Google Scholar 

  • J. Offord W.A. Catterall (1989) ArticleTitleElectrical activity, cAMP, and cytosolic calcium regulate mRNA encoding sodium channel alpha subunits in rat muscle cells Neuron 2 1447–1452 Occurrence Handle10.1016/0896-6273(89)90190-6 Occurrence Handle1:CAS:528:DyaK3cXltFaqs7k%3D Occurrence Handle2560642

    Article  CAS  PubMed  Google Scholar 

  • N. Oka K. Asai R.K. Kudej J.G. Edwards Y. Toya C. Schwencke D.E. Vatner S.F. Vatner Y. Ishikawa (1997) ArticleTitleDownregulation of caveolin by chronic β-adrenergic receptor stimulation in mice Am. J. Physiol 273 C1957–C1962 Occurrence Handle1:CAS:528:DyaK1cXjvFWisQ%3D%3D Occurrence Handle9435501

    CAS  PubMed  Google Scholar 

  • M. Pragnell M. Waard ParticleDe Y. Mori T. Tanabe T.P. Snutch K.P. Campbell (1994) ArticleTitleCalcium channel β-subunit binds to a conserved motif in the I–II cytoplasmic linker of the α1-subunit Nature 368 67–70 Occurrence Handle10.1038/368067a0 Occurrence Handle1:CAS:528:DyaK2cXis1eqsbY%3D Occurrence Handle7509046

    Article  CAS  PubMed  Google Scholar 

  • M. Rechsteiner S.W. Rogers (1996) ArticleTitlePEST sequences and regulation by proteolysis Trends Biochem. Sci 21 267–271 Occurrence Handle10.1016/0968-0004(96)10031-1 Occurrence Handle1:CAS:528:DyaK28XksVymtr8%3D Occurrence Handle8755249

    Article  CAS  PubMed  Google Scholar 

  • C. Romanin P. Grosswagen H. Schindler (1991) ArticleTitleCalpastatin and nucleotides stabilize cardiac calcium channel activity in excised patches Pfluegers Arch 418 86–92 Occurrence Handle10.1007/BF00370456 Occurrence Handle1:CAS:528:DyaK3MXisVaqsbk%3D

    Article  CAS  Google Scholar 

  • J.A. Sánchez E. Stefani (1983) ArticleTitleKinetic properties of calcium channels of twitch muscle fibres of the frog J. Physiol 337 1–17 Occurrence Handle6308234

    PubMed  Google Scholar 

  • S. Shiraishi I. Shibuya Y. Uezono H. Yokoo Y. Toyohira R. Yamamoto T. Yanagita H. Kobayashi A. Wada (2001) ArticleTitleHeterogeneous increases of cytoplasmic calcium: distinct effects on down-regulation of cell surface sodium channels and sodium channel subunit mRNA levels Br. J. Pharmacol 132 1455–1466 Occurrence Handle10.1038/sj.bjp.0703960 Occurrence Handle1:CAS:528:DC%2BD3MXivVahs7k%3D Occurrence Handle11264239

    Article  CAS  PubMed  Google Scholar 

  • H. Sorimachi K. Suzuki (2001) ArticleTitleThe structure of calpain J. Biochem. (Tokyo) 129 653–664 Occurrence Handle1:CAS:528:DC%2BD3MXltVWltLs%3D

    CAS  Google Scholar 

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Acknowledgments

The authors are most grateful to Dr. Philip Palade for providing advice on RNAse protection assays. We thank Mr. Oscar Ramírez for excellent technical assistance and Ms. Susana Zamudio for excellent secretarial work. This work was supported by CONACyT grant #32055N to JS, grant 37356-N # to MG and by NIH grant (DHHS) 1 R01 HL63903-01A1 and 18 (UC-MEX).

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  1. Department of Pharmacology, Cinvestav, A.P.14-740, México, D.F., 07300

    E. Carrillo, J.M. Galindo, M.C. García & J.A. Sánchez

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  1. E. Carrillo
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  2. J.M. Galindo
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  3. M.C. García
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  4. J.A. Sánchez
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Correspondence to J.A. Sánchez.

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Carrillo, E., Galindo, J., García, M. et al. Regulation of Muscle Cav1.1 Channels by Long-term Depolarization Involves Proteolysis of the α1s Subunit. J Membrane Biol 199, 155–161 (2004). https://doi.org/10.1007/s00232-004-0683-x

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  • DOI: https://doi.org/10.1007/s00232-004-0683-x

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