Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 1995 Feb;68(2):471–482. doi: 10.1016/S0006-3495(95)80208-0

Physiological differences between the alpha and beta ryanodine receptors of fish skeletal muscle.

J O'Brien 1, H H Valdivia 1, B A Block 1
PMCID: PMC1281711  PMID: 7696500

Abstract

Two isoforms of the sarcoplasmic reticulum Ca2+ release channel (ryanodine receptor or RYR) are expressed together in the skeletal muscles of most vertebrates. We have studied physiological properties of the two isoforms (alpha and beta) by comparing SR preparations from specialized fish muscles that express the alpha isoform alone to preparations from muscles containing both alpha and beta. Regulation of channel activity was assessed through [3H]ryanodine binding and reconstitution into planar lipid bilayers. Distinct differences were observed in the calcium-activation and -inactivation properties of the two isoforms. The fish alpha isoform, expressed alone in extraocular muscles, closely resembled the rabbit skeletal muscle RYR. Maximum [3H]ryanodine binding and maximum open probability (Po) of the alpha RYR were achieved from 1 to 10 microM free Ca2+. Millimolar Ca2+ reduced [3H]ryanodine binding and Po close to zero. The beta isoform more closely resembled the fish cardiac RYR in Ca2+ activation of [3H]ryanodine binding. The most prominent difference of the beta and cardiac isoforms from the alpha isoform was the lack of inactivation of [3H]ryanodine binding and Po by millimolar free Ca2+. Differences in activation of [3H]ryanodine binding by adenine nucleotides and inhibition by Mg2+ suggest that the beta and cardiac RYRs are not identical, however. [3H]ryanodine binding by the alpha RYR was selectively inhibited by 100 microM tetracaine, whereas cardiac and beta RYRs were much less affected. Tetracaine can thus be used to separate the properties of the alpha and beta RYRs in preparations in which both are present. The distinct physiological properties of the alpha and beta RYRs that are present together in most vertebrate muscles support models of EC coupling incorporating both directly coupled and Ca(2+)-coupled channels within a single triad junction.

Full text

PDF
471

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Airey J. A., Beck C. F., Murakami K., Tanksley S. J., Deerinck T. J., Ellisman M. H., Sutko J. L. Identification and localization of two triad junctional foot protein isoforms in mature avian fast twitch skeletal muscle. J Biol Chem. 1990 Aug 25;265(24):14187–14194. [PubMed] [Google Scholar]
  2. Airey J. A., Grinsell M. M., Jones L. R., Sutko J. L., Witcher D. Three ryanodine receptor isoforms exist in avian striated muscles. Biochemistry. 1993 Jun 8;32(22):5739–5745. doi: 10.1021/bi00073a003. [DOI] [PubMed] [Google Scholar]
  3. Almers W., Best P. M. Effects of tetracaine on displacement currents and contraction of frog skeletal muscle. J Physiol. 1976 Nov;262(3):583–611. doi: 10.1113/jphysiol.1976.sp011611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anderson K., Lai F. A., Liu Q. Y., Rousseau E., Erickson H. P., Meissner G. Structural and functional characterization of the purified cardiac ryanodine receptor-Ca2+ release channel complex. J Biol Chem. 1989 Jan 15;264(2):1329–1335. [PubMed] [Google Scholar]
  5. Baylor S. M., Hollingworth S. Fura-2 calcium transients in frog skeletal muscle fibres. J Physiol. 1988 Sep;403:151–192. doi: 10.1113/jphysiol.1988.sp017244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Block B. A., Imagawa T., Campbell K. P., Franzini-Armstrong C. Structural evidence for direct interaction between the molecular components of the transverse tubule/sarcoplasmic reticulum junction in skeletal muscle. J Cell Biol. 1988 Dec;107(6 Pt 2):2587–2600. doi: 10.1083/jcb.107.6.2587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bull R., Marengo J. J. Sarcoplasmic reticulum release channels from frog skeletal muscle display two types of calcium dependence. FEBS Lett. 1993 Oct 4;331(3):223–227. doi: 10.1016/0014-5793(93)80341-q. [DOI] [PubMed] [Google Scholar]
  8. Chu A., Díaz-Muñoz M., Hawkes M. J., Brush K., Hamilton S. L. Ryanodine as a probe for the functional state of the skeletal muscle sarcoplasmic reticulum calcium release channel. Mol Pharmacol. 1990 May;37(5):735–741. [PubMed] [Google Scholar]
  9. Chu A., Fill M., Stefani E., Entman M. L. Cytoplasmic Ca2+ does not inhibit the cardiac muscle sarcoplasmic reticulum ryanodine receptor Ca2+ channel, although Ca(2+)-induced Ca2+ inactivation of Ca2+ release is observed in native vesicles. J Membr Biol. 1993 Jul;135(1):49–59. doi: 10.1007/BF00234651. [DOI] [PubMed] [Google Scholar]
  10. Csernoch L., Huang C. L., Szucs G., Kovacs L. Differential effects of tetracaine on charge movements and Ca2+ signals in frog skeletal muscle. J Gen Physiol. 1988 Nov;92(5):601–612. doi: 10.1085/jgp.92.5.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Csernoch L., Jacquemond V., Schneider M. F. Microinjection of strong calcium buffers suppresses the peak of calcium release during depolarization in frog skeletal muscle fibers. J Gen Physiol. 1993 Feb;101(2):297–333. doi: 10.1085/jgp.101.2.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fabiato A., Fabiato F. Calculator programs for computing the composition of the solutions containing multiple metals and ligands used for experiments in skinned muscle cells. J Physiol (Paris) 1979;75(5):463–505. [PubMed] [Google Scholar]
  13. Hakamata Y., Nakai J., Takeshima H., Imoto K. Primary structure and distribution of a novel ryanodine receptor/calcium release channel from rabbit brain. FEBS Lett. 1992 Nov 9;312(2-3):229–235. doi: 10.1016/0014-5793(92)80941-9. [DOI] [PubMed] [Google Scholar]
  14. Hollingworth S., Harkins A. B., Kurebayashi N., Konishi M., Baylor S. M. Excitation-contraction coupling in intact frog skeletal muscle fibers injected with mmolar concentrations of fura-2. Biophys J. 1992 Jul;63(1):224–234. doi: 10.1016/S0006-3495(92)81599-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Inui M., Saito A., Fleischer S. Isolation of the ryanodine receptor from cardiac sarcoplasmic reticulum and identity with the feet structures. J Biol Chem. 1987 Nov 15;262(32):15637–15642. [PubMed] [Google Scholar]
  16. Inui M., Saito A., Fleischer S. Purification of the ryanodine receptor and identity with feet structures of junctional terminal cisternae of sarcoplasmic reticulum from fast skeletal muscle. J Biol Chem. 1987 Feb 5;262(4):1740–1747. [PubMed] [Google Scholar]
  17. Jacquemond V., Csernoch L., Klein M. G., Schneider M. F. Voltage-gated and calcium-gated calcium release during depolarization of skeletal muscle fibers. Biophys J. 1991 Oct;60(4):867–873. doi: 10.1016/S0006-3495(91)82120-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jong D. S., Pape P. C., Chandler W. K., Baylor S. M. Reduction of calcium inactivation of sarcoplasmic reticulum calcium release by fura-2 in voltage-clamped cut twitch fibers from frog muscle. J Gen Physiol. 1993 Aug;102(2):333–370. doi: 10.1085/jgp.102.2.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kaplan R. S., Pedersen P. L. Determination of microgram quantities of protein in the presence of milligram levels of lipid with amido black 10B. Anal Biochem. 1985 Oct;150(1):97–104. doi: 10.1016/0003-2697(85)90445-2. [DOI] [PubMed] [Google Scholar]
  20. Kuwajima G., Futatsugi A., Niinobe M., Nakanishi S., Mikoshiba K. Two types of ryanodine receptors in mouse brain: skeletal muscle type exclusively in Purkinje cells and cardiac muscle type in various neurons. Neuron. 1992 Dec;9(6):1133–1142. doi: 10.1016/0896-6273(92)90071-k. [DOI] [PubMed] [Google Scholar]
  21. Lai F. A., Erickson H. P., Rousseau E., Liu Q. Y., Meissner G. Purification and reconstitution of the calcium release channel from skeletal muscle. Nature. 1988 Jan 28;331(6154):315–319. doi: 10.1038/331315a0. [DOI] [PubMed] [Google Scholar]
  22. Lai F. A., Liu Q. Y., Xu L., el-Hashem A., Kramarcy N. R., Sealock R., Meissner G. Amphibian ryanodine receptor isoforms are related to those of mammalian skeletal or cardiac muscle. Am J Physiol. 1992 Aug;263(2 Pt 1):C365–C372. doi: 10.1152/ajpcell.1992.263.2.C365. [DOI] [PubMed] [Google Scholar]
  23. Lai F. A., Misra M., Xu L., Smith H. A., Meissner G. The ryanodine receptor-Ca2+ release channel complex of skeletal muscle sarcoplasmic reticulum. Evidence for a cooperatively coupled, negatively charged homotetramer. J Biol Chem. 1989 Oct 5;264(28):16776–16785. [PubMed] [Google Scholar]
  24. Liu Q. Y., Lai F. A., Rousseau E., Jones R. V., Meissner G. Multiple conductance states of the purified calcium release channel complex from skeletal sarcoplasmic reticulum. Biophys J. 1989 Mar;55(3):415–424. doi: 10.1016/S0006-3495(89)82835-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Marks A. R., Tempst P., Hwang K. S., Taubman M. B., Inui M., Chadwick C., Fleischer S., Nadal-Ginard B. Molecular cloning and characterization of the ryanodine receptor/junctional channel complex cDNA from skeletal muscle sarcoplasmic reticulum. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8683–8687. doi: 10.1073/pnas.86.22.8683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Meissner G., Darling E., Eveleth J. Kinetics of rapid Ca2+ release by sarcoplasmic reticulum. Effects of Ca2+, Mg2+, and adenine nucleotides. Biochemistry. 1986 Jan 14;25(1):236–244. doi: 10.1021/bi00349a033. [DOI] [PubMed] [Google Scholar]
  27. Meissner G. Ryanodine activation and inhibition of the Ca2+ release channel of sarcoplasmic reticulum. J Biol Chem. 1986 May 15;261(14):6300–6306. [PubMed] [Google Scholar]
  28. Meissner G. Ryanodine receptor/Ca2+ release channels and their regulation by endogenous effectors. Annu Rev Physiol. 1994;56:485–508. doi: 10.1146/annurev.ph.56.030194.002413. [DOI] [PubMed] [Google Scholar]
  29. Michalak M., Dupraz P., Shoshan-Barmatz V. Ryanodine binding to sarcoplasmic reticulum membrane; comparison between cardiac and skeletal muscle. Biochim Biophys Acta. 1988 Apr 22;939(3):587–594. doi: 10.1016/0005-2736(88)90106-x. [DOI] [PubMed] [Google Scholar]
  30. Murayama T., Ogawa Y. Purification and characterization of two ryanodine-binding protein isoforms from sarcoplasmic reticulum of bullfrog skeletal muscle. J Biochem. 1992 Oct;112(4):514–522. doi: 10.1093/oxfordjournals.jbchem.a123931. [DOI] [PubMed] [Google Scholar]
  31. Nakai J., Imagawa T., Hakamat Y., Shigekawa M., Takeshima H., Numa S. Primary structure and functional expression from cDNA of the cardiac ryanodine receptor/calcium release channel. FEBS Lett. 1990 Oct 1;271(1-2):169–177. doi: 10.1016/0014-5793(90)80399-4. [DOI] [PubMed] [Google Scholar]
  32. O'Brien J., Meissner G., Block B. A. The fastest contracting muscles of nonmammalian vertebrates express only one isoform of the ryanodine receptor. Biophys J. 1993 Dec;65(6):2418–2427. doi: 10.1016/S0006-3495(93)81303-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Olivares E. B., Tanksley S. J., Airey J. A., Beck C. F., Ouyang Y., Deerinck T. J., Ellisman M. H., Sutko J. L. Nonmammalian vertebrate skeletal muscles express two triad junctional foot protein isoforms. Biophys J. 1991 Jun;59(6):1153–1163. doi: 10.1016/S0006-3495(91)82331-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Otsu K., Willard H. F., Khanna V. K., Zorzato F., Green N. M., MacLennan D. H. Molecular cloning of cDNA encoding the Ca2+ release channel (ryanodine receptor) of rabbit cardiac muscle sarcoplasmic reticulum. J Biol Chem. 1990 Aug 15;265(23):13472–13483. [PubMed] [Google Scholar]
  35. Ouyang Y., Deerinck T. J., Walton P. D., Airey J. A., Sutko J. L., Ellisman M. H. Distribution of ryanodine receptors in the chicken central nervous system. Brain Res. 1993 Aug 27;620(2):269–280. doi: 10.1016/0006-8993(93)90165-j. [DOI] [PubMed] [Google Scholar]
  36. Oyamada H., Murayama T., Takagi T., Iino M., Iwabe N., Miyata T., Ogawa Y., Endo M. Primary structure and distribution of ryanodine-binding protein isoforms of the bullfrog skeletal muscle. J Biol Chem. 1994 Jun 24;269(25):17206–17214. [PubMed] [Google Scholar]
  37. Pape P. C., Jong D. S., Chandler W. K., Baylor S. M. Effect of fura-2 on action potential-stimulated calcium release in cut twitch fibers from frog muscle. J Gen Physiol. 1993 Aug;102(2):295–332. doi: 10.1085/jgp.102.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Pessah I. N., Stambuk R. A., Casida J. E. Ca2+-activated ryanodine binding: mechanisms of sensitivity and intensity modulation by Mg2+, caffeine, and adenine nucleotides. Mol Pharmacol. 1987 Mar;31(3):232–238. [PubMed] [Google Scholar]
  39. Pessah I. N., Waterhouse A. L., Casida J. E. The calcium-ryanodine receptor complex of skeletal and cardiac muscle. Biochem Biophys Res Commun. 1985 Apr 16;128(1):449–456. doi: 10.1016/0006-291x(85)91699-7. [DOI] [PubMed] [Google Scholar]
  40. Pizarro G., Csernoch L., Uribe I., Ríos E. Differential effects of tetracaine on two kinetic components of calcium release in frog skeletal muscle fibres. J Physiol. 1992 Nov;457:525–538. doi: 10.1113/jphysiol.1992.sp019392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rousseau E., Smith J. S., Henderson J. S., Meissner G. Single channel and 45Ca2+ flux measurements of the cardiac sarcoplasmic reticulum calcium channel. Biophys J. 1986 Nov;50(5):1009–1014. doi: 10.1016/S0006-3495(86)83543-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ríos E., Karhanek M., Ma J., González A. An allosteric model of the molecular interactions of excitation-contraction coupling in skeletal muscle. J Gen Physiol. 1993 Sep;102(3):449–481. doi: 10.1085/jgp.102.3.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ríos E., Ma J. J., González A. The mechanical hypothesis of excitation-contraction (EC) coupling in skeletal muscle. J Muscle Res Cell Motil. 1991 Apr;12(2):127–135. doi: 10.1007/BF01774031. [DOI] [PubMed] [Google Scholar]
  44. Schneider M. F., Chandler W. K. Voltage dependent charge movement of skeletal muscle: a possible step in excitation-contraction coupling. Nature. 1973 Mar 23;242(5395):244–246. doi: 10.1038/242244a0. [DOI] [PubMed] [Google Scholar]
  45. Schneider M. F., Simon B. J. Inactivation of calcium release from the sarcoplasmic reticulum in frog skeletal muscle. J Physiol. 1988 Nov;405:727–745. doi: 10.1113/jphysiol.1988.sp017358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Simon B. J., Schneider M. F. Time course of activation of calcium release from sarcoplasmic reticulum in skeletal muscle. Biophys J. 1988 Dec;54(6):1159–1163. doi: 10.1016/S0006-3495(88)83050-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Smith J. S., Coronado R., Meissner G. Single channel measurements of the calcium release channel from skeletal muscle sarcoplasmic reticulum. Activation by Ca2+ and ATP and modulation by Mg2+. J Gen Physiol. 1986 Nov;88(5):573–588. doi: 10.1085/jgp.88.5.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Smith J. S., Imagawa T., Ma J., Fill M., Campbell K. P., Coronado R. Purified ryanodine receptor from rabbit skeletal muscle is the calcium-release channel of sarcoplasmic reticulum. J Gen Physiol. 1988 Jul;92(1):1–26. doi: 10.1085/jgp.92.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Stern M. D. Theory of excitation-contraction coupling in cardiac muscle. Biophys J. 1992 Aug;63(2):497–517. doi: 10.1016/S0006-3495(92)81615-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Strand M. A., Louis C. F., Mickelson J. R. Phosphorylation of the porcine skeletal and cardiac muscle sarcoplasmic reticulum ryanodine receptor. Biochim Biophys Acta. 1993 Feb 17;1175(3):319–326. doi: 10.1016/0167-4889(93)90224-d. [DOI] [PubMed] [Google Scholar]
  51. Takeshima H., Nishimura S., Nishi M., Ikeda M., Sugimoto T. A brain-specific transcript from the 3'-terminal region of the skeletal muscle ryanodine receptor gene. FEBS Lett. 1993 May 10;322(2):105–110. doi: 10.1016/0014-5793(93)81547-d. [DOI] [PubMed] [Google Scholar]
  52. Tinker A., Lindsay A. R., Williams A. J. A model for ionic conduction in the ryanodine receptor channel of sheep cardiac muscle sarcoplasmic reticulum. J Gen Physiol. 1992 Sep;100(3):495–517. doi: 10.1085/jgp.100.3.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Valdivia H. H., Kirby M. S., Lederer W. J., Coronado R. Scorpion toxins targeted against the sarcoplasmic reticulum Ca(2+)-release channel of skeletal and cardiac muscle. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12185–12189. doi: 10.1073/pnas.89.24.12185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Witcher D. R., Kovacs R. J., Schulman H., Cefali D. C., Jones L. R. Unique phosphorylation site on the cardiac ryanodine receptor regulates calcium channel activity. J Biol Chem. 1991 Jun 15;266(17):11144–11152. [PubMed] [Google Scholar]
  55. Xu L., Jones R., Meissner G. Effects of local anesthetics on single channel behavior of skeletal muscle calcium release channel. J Gen Physiol. 1993 Feb;101(2):207–233. doi: 10.1085/jgp.101.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES