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. 1995 Dec 15;489(Pt 3):791–803. doi: 10.1113/jphysiol.1995.sp021092

Synthesis and release of an acetylcholine-like compound by human myoblasts and myotubes.

M Hamann 1, M C Chamoin 1, P Portalier 1, L Bernheim 1, A Baroffio 1, H Widmer 1, C R Bader 1, J P Ternaux 1
PMCID: PMC1156848  PMID: 8788943

Abstract

1. Exogenously applied acetylcholine (ACh) is a modulator of human myoblast fusion. Using a chemiluminescent method, we examined whether an endogenous ACh-like compound (ACh-lc) was present in, and released by, pure human myogenic cells. 2. Single, freshly isolated satellite cells and proliferating myoblasts contained 15 and 0.5 fmol ACh-lc, respectively. Cultured myotubes contained ACh-lc as well. Also, ACh-like immunoreactivity was detected in all myogenic cells. 3. Part of the ACh-lc was synthesized by choline acetyltransferase (ChAT), as indicated by the reduction of ACh-lc content when bromoACh was present in the culture medium, and by direct measurements of ChAT activity. Also, ChAT-like immunoreactivity was observed in all myogenic cells. 4. Myoblasts and myotubes released ACh-lc spontaneously by a partially Ca(2+)-dependent mechanism. 5. The application by microperfusion of medium conditioned beforehand by myoblasts (thus presumably containing ACh-lc) onto a voltage-clamped myotube induced inward currents resembling ACh-induced currents in their kinetics, reversal potential, and sensitivity to nicotinic antagonists. 6. In vitro, the spontaneously released ACh-lc promoted myoblast fusion but only in the presence of an anticholinesterase. 7. Our observations indicate that human myogenic cells synthesize and release an ACh-lc and thereby promote the fusion process that occurs in muscle during growth or regeneration.

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Selected References

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

  1. Bacou F., Vigneron P. Neural influence on the expression of acetylcholinesterase molecular forms in fast and slow rabbit skeletal muscles. Dev Biol. 1991 Jun;145(2):356–366. doi: 10.1016/0012-1606(91)90134-o. [DOI] [PubMed] [Google Scholar]
  2. Bader C. R., Bertrand D., Cooper E., Mauro A. Membrane currents of rat satellite cells attached to intact skeletal muscle fibers. Neuron. 1988 May;1(3):237–240. doi: 10.1016/0896-6273(88)90144-4. [DOI] [PubMed] [Google Scholar]
  3. Bakry N. M., el-Rashidy A. H., Eldefrawi A. T., Eldefrawi M. E. Direct actions of organophosphate anticholinesterases on nicotinic and muscarinic acetylcholine receptors. J Biochem Toxicol. 1988 Winter;3:235–259. doi: 10.1002/jbt.2570030404. [DOI] [PubMed] [Google Scholar]
  4. Baroffio A., Aubry J. P., Kaelin A., Krause R. M., Hamann M., Bader C. R. Purification of human muscle satellite cells by flow cytometry. Muscle Nerve. 1993 May;16(5):498–505. doi: 10.1002/mus.880160511. [DOI] [PubMed] [Google Scholar]
  5. Bertrand D., Bader C. R. DATAC: a multipurpose biological data analysis program based on a mathematical interpreter. Int J Biomed Comput. 1986 May;18(3-4):193–202. doi: 10.1016/0020-7101(86)90016-4. [DOI] [PubMed] [Google Scholar]
  6. Bowman W. C., Marshall I. G., Gibb A. J., Harborne A. J. Feedback control of transmitter release at the neuromuscular junction. Trends Pharmacol Sci. 1988 Jan;9(1):16–20. doi: 10.1016/0165-6147(88)90236-2. [DOI] [PubMed] [Google Scholar]
  7. Bray J. J., Forrest J. W., Hubbard J. I. Evidence for the role of non-quantal acetylcholine in the maintenance of the membrane potential of rat skeletal muscle. J Physiol. 1982 May;326:285–296. doi: 10.1113/jphysiol.1982.sp014192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Buchanan J., Sun Y. A., Poo M. M. Studies of nerve-muscle interactions in Xenopus cell culture: fine structure of early functional contacts. J Neurosci. 1989 May;9(5):1540–1554. doi: 10.1523/JNEUROSCI.09-05-01540.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Campion D. R. The muscle satellite cell: a review. Int Rev Cytol. 1984;87:225–251. doi: 10.1016/s0074-7696(08)62444-4. [DOI] [PubMed] [Google Scholar]
  10. Dale H. H., Feldberg W., Vogt M. Release of acetylcholine at voluntary motor nerve endings. J Physiol. 1936 May 4;86(4):353–380. doi: 10.1113/jphysiol.1936.sp003371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dan Y., Poo M. M. Quantal transmitter secretion from myocytes loaded with acetylcholine. Nature. 1992 Oct 22;359(6397):733–736. doi: 10.1038/359733a0. [DOI] [PubMed] [Google Scholar]
  12. Dennis M. J., Miledi R. Electrically induced release of acetylcholine from denervated Schwann cells. J Physiol. 1974 Mar;237(2):431–452. doi: 10.1113/jphysiol.1974.sp010490. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dlouhá H., Teisinger J., Vyskocil F. Activation of membrane Na+/K+-ATPase of mouse skeletal muscle by acetylcholine and its inhibition by alpha-bungarotoxin, curare and atropine. Pflugers Arch. 1979 May 15;380(1):101–104. doi: 10.1007/BF00582620. [DOI] [PubMed] [Google Scholar]
  14. Elson H. F., Gentry M. K., Doctor B. P. Membrane-bound acetylcholinesterase: an early differentiation marker for skeletal myoblasts. Biochim Biophys Acta. 1992 Dec 8;1156(1):78–84. doi: 10.1016/0304-4165(92)90099-g. [DOI] [PubMed] [Google Scholar]
  15. Entwistle A., Zalin R. J., Bevan S., Warner A. E. The control of chick myoblast fusion by ion channels operated by prostaglandins and acetylcholine. J Cell Biol. 1988 May;106(5):1693–1702. doi: 10.1083/jcb.106.5.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Entwistle A., Zalin R. J., Warner A. E., Bevan S. A role for acetylcholine receptors in the fusion of chick myoblasts. J Cell Biol. 1988 May;106(5):1703–1712. doi: 10.1083/jcb.106.5.1703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Eusebi F., Molinaro M. Acetylcholine sensitivity in replicating satellite cells. Muscle Nerve. 1984 Jul-Aug;7(6):488–492. doi: 10.1002/mus.880070613. [DOI] [PubMed] [Google Scholar]
  18. Fonnum F. Radiochemical micro assays for the determination of choline acetyltransferase and acetylcholinesterase activities. Biochem J. 1969 Nov;115(3):465–472. doi: 10.1042/bj1150465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Geffard M., McRae-Degueurce A., Souan M. L. Immunocytochemical detection of acetylcholine in the rat central nervous system. Science. 1985 Jul 5;229(4708):77–79. doi: 10.1126/science.3892687. [DOI] [PubMed] [Google Scholar]
  20. HEBB C. O., KRNJEVIC K., SILVER A. ACETYLCHOLINE AND CHOLINE ACETYLTRANSFERASE IN THE DIAPHRAGM OF THE RAT. J Physiol. 1964 Jun;171:504–513. doi: 10.1113/jphysiol.1964.sp007393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hamann M., Widmer H., Baroffio A., Aubry J. P., Krause R. M., Kaelin A., Bader C. R. Sodium and potassium currents in freshly isolated and in proliferating human muscle satellite cells. J Physiol. 1994 Mar 1;475(2):305–317. doi: 10.1113/jphysiol.1994.sp020071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  23. Hebb C. O. Acetylcholine content of the rabbit plantaris muscle after denervation. J Physiol. 1962 Sep;163(2):294–306. doi: 10.1113/jphysiol.1962.sp006975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Israel M., Dunant Y., Manaranche R. The present status of the vesicular hypothesis. Prog Neurobiol. 1979;13(3):237–275. doi: 10.1016/0301-0082(79)90017-0. [DOI] [PubMed] [Google Scholar]
  25. Katz B., Miledi R. Transmitter leakage from motor nerve endings. Proc R Soc Lond B Biol Sci. 1977 Feb 11;196(1122):59–72. doi: 10.1098/rspb.1977.0029. [DOI] [PubMed] [Google Scholar]
  26. Krause R. M., Hamann M., Bader C. R., Liu J. H., Baroffio A., Bernheim L. Activation of nicotinic acetylcholine receptors increases the rate of fusion of cultured human myoblasts. J Physiol. 1995 Dec 15;489(Pt 3):779–790. doi: 10.1113/jphysiol.1995.sp021091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  28. Lankford K. L., DeMello F. G., Klein W. L. D1-type dopamine receptors inhibit growth cone motility in cultured retina neurons: evidence that neurotransmitters act as morphogenic growth regulators in the developing central nervous system. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2839–2843. doi: 10.1073/pnas.85.8.2839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lipton S. A. Spontaneous release of acetylcholine affects the physiological nicotinic responses of rat retinal ganglion cells in culture. J Neurosci. 1988 Oct;8(10):3857–3868. doi: 10.1523/JNEUROSCI.08-10-03857.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Mattson M. P. Neurotransmitters in the regulation of neuronal cytoarchitecture. Brain Res. 1988 Apr-Jun;472(2):179–212. doi: 10.1016/0165-0173(88)90020-3. [DOI] [PubMed] [Google Scholar]
  31. McCobb D. P., Cohan C. S., Connor J. A., Kater S. B. Interactive effects of serotonin and acetylcholine on neurite elongation. Neuron. 1988 Jul;1(5):377–385. doi: 10.1016/0896-6273(88)90187-0. [DOI] [PubMed] [Google Scholar]
  32. Miledi R., Molenaar P. C., Polak R. L. An analysis of acetylcholine in frog muscle by mass fragmentography. Proc R Soc Lond B Biol Sci. 1977 Jun 15;197(1128):285–297. doi: 10.1098/rspb.1977.0071. [DOI] [PubMed] [Google Scholar]
  33. Miledi R., Molenaar P. C., Polak R. L., Tas J. W., van der Laaken T. Neural and non-neural acetylcholine in the rat diaphragm. Proc R Soc Lond B Biol Sci. 1982 Jan 22;214(1195):153–168. doi: 10.1098/rspb.1982.0002. [DOI] [PubMed] [Google Scholar]
  34. Mitchell J. F., Silver A. The spontaneous release of acetylcholine from the denervated hemidiaphragm of the rat. J Physiol. 1963 Jan;165(1):117–129. doi: 10.1113/jphysiol.1963.sp007046. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Molenaar P. C., Oen B. S., Polak R. L., van der Laaken A. L. Surplus acetylcholine and acetylcholine release in the rat diaphragm. J Physiol. 1987 Apr;385:147–167. doi: 10.1113/jphysiol.1987.sp016489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Palouzier-Paulignan B., Chamoin M.-C., Ternaux J.-P. Somatic Acetylcholine Release in Rabbit Nodose Ganglion. Eur J Neurosci. 1992 Oct;4(11):1123–1129. doi: 10.1111/j.1460-9568.1992.tb00139.x. [DOI] [PubMed] [Google Scholar]
  37. Pestronk A., Drachman D. B., Stanley E. F., Price D. L., Griffin J. W. Cholinergic transmission regulates extrajunctional acetylcholine receptors. Exp Neurol. 1980 Dec;70(3):690–696. doi: 10.1016/0014-4886(80)90193-4. [DOI] [PubMed] [Google Scholar]
  38. Schubert D., Heinemann S., Carlisle W., Tarikas H., Kimes B., Patrick J., Steinbach J. H., Culp W., Brandt B. L. Clonal cell lines from the rat central nervous system. Nature. 1974 May 17;249(454):224–227. doi: 10.1038/249224a0. [DOI] [PubMed] [Google Scholar]
  39. Siegelbaum S. A., Trautmann A., Koenig J. Single acetylcholine-activated channel currents in developing muscle cells. Dev Biol. 1984 Aug;104(2):366–379. doi: 10.1016/0012-1606(84)90092-7. [DOI] [PubMed] [Google Scholar]
  40. Ternaux J. P., Portalier P. Influence of tongue myoblasts on rat dissociated hypoglossal motoneurons in culture. Int J Dev Neurosci. 1993 Feb;11(1):33–48. doi: 10.1016/0736-5748(93)90033-a. [DOI] [PubMed] [Google Scholar]
  41. Tucek S. The synthesis of acetylcholine in skeletal muscles of the rat. J Physiol. 1982 Jan;322:53–69. doi: 10.1113/jphysiol.1982.sp014022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. White H. L., Wu J. C. Choline and carnitine acetyltransferases of heart. Biochemistry. 1973 Feb 27;12(5):841–846. doi: 10.1021/bi00729a009. [DOI] [PubMed] [Google Scholar]
  43. Widmer H., Hamann M., Baroffio A., Bijlenga P., Bader C. R. Expression of a voltage-dependent potassium current precedes fusion of human muscle satellite cells (myoblasts). J Cell Physiol. 1995 Jan;162(1):52–63. doi: 10.1002/jcp.1041620108. [DOI] [PubMed] [Google Scholar]

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