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. 1987 Jan 1;104(1):87–95. doi: 10.1083/jcb.104.1.87

Acetylcholine receptor clustering and nuclear movement in muscle fibers in culture

PMCID: PMC2117039  PMID: 3793762

Abstract

We have studied the formation of acetylcholine receptor (AChR) clusters and the behavior of myonuclei in rat and chick skeletal muscle cells grown in cell culture. These cells were treated with a factor derived from Torpedo electric extracellular matrix, which causes a large increase in their number of AChR clusters. We found that these clusters were located preferentially in membrane regions above myonuclei. This cluster-nucleus colocalization is explained by our finding that most of the nuclei near clusters remain relatively stationary, while most of those away from clusters are able to translocate throughout the myotube. In some cases, clusters clearly formed first, then nuclei migrated underneath and became immobilized. If clustered AChRs later dispersed, their associated nuclei resumed moving. These results suggest that AChR clustering initiates an extensive cytoskeletal rearrangement that causes the subcluster localization of organelles, potentially providing a stable source of newly synthesized AChRs for insertion into the cluster.

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Selected References

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  1. Anderson M. J., Cohen M. W. Nerve-induced and spontaneous redistribution of acetylcholine receptors on cultured muscle cells. J Physiol. 1977 Jul;268(3):757–773. doi: 10.1113/jphysiol.1977.sp011880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson M. J., Cohen M. W., Zorychta E. Effects of innervation on the distribution of acetylcholine receptors on cultured muscle cells. J Physiol. 1977 Jul;268(3):731–756. doi: 10.1113/jphysiol.1977.sp011879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anthony D. T., Schuetze S. M., Rubin L. L. Transformation by Rous sarcoma virus prevents acetylcholine receptor clustering on cultured chicken muscle fibers. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2265–2269. doi: 10.1073/pnas.81.7.2265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bloch R. J. Dispersal and reformation of acetylcholine receptor clusters of cultured rat myotubes treated with inhibitors of energy metabolism. J Cell Biol. 1979 Sep;82(3):626–643. doi: 10.1083/jcb.82.3.626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bloch R. J., Hall Z. W. Cytoskeletal components of the vertebrate neuromuscular junction: vinculin, alpha-actinin, and filamin. J Cell Biol. 1983 Jul;97(1):217–223. doi: 10.1083/jcb.97.1.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burden S. J., DePalma R. L., Gottesman G. S. Crosslinking of proteins in acetylcholine receptor-rich membranes: association between the beta-subunit and the 43 kd subsynaptic protein. Cell. 1983 Dec;35(3 Pt 2):687–692. doi: 10.1016/0092-8674(83)90101-0. [DOI] [PubMed] [Google Scholar]
  7. Burden S. J., Sargent P. B., McMahan U. J. Acetylcholine receptors in regenerating muscle accumulate at original synaptic sites in the absence of the nerve. J Cell Biol. 1979 Aug;82(2):412–425. doi: 10.1083/jcb.82.2.412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burden S. Identification of an intracellular postsynaptic antigen at the frog neuromuscular junction. J Cell Biol. 1982 Sep;94(3):521–530. doi: 10.1083/jcb.94.3.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Bursztajn S., Fischbach G. D. Evidence that coated vesicles transport acetylcholine receptors to the surface membrane of chick myotubes. J Cell Biol. 1984 Feb;98(2):498–506. doi: 10.1083/jcb.98.2.498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Connolly J. A., Graham A. J. Actin filaments and acetylcholine receptor clusters in embryonic chick myotubes. Eur J Cell Biol. 1985 May;37:191–195. [PubMed] [Google Scholar]
  11. Connolly J. A. Role of the cytoskeleton in the formation, stabilization, and removal of acetylcholine receptor clusters in cultured muscle cells. J Cell Biol. 1984 Jul;99(1 Pt 1):148–154. doi: 10.1083/jcb.99.1.148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dennis M. J. Development of the neuromuscular junction: inductive interactions between cells. Annu Rev Neurosci. 1981;4:43–68. doi: 10.1146/annurev.ne.04.030181.000355. [DOI] [PubMed] [Google Scholar]
  13. Devreotes P. N., Fambrough D. M. Acetylcholine receptor turnover in membranes of developing muscle fibers. J Cell Biol. 1975 May;65(2):335–358. doi: 10.1083/jcb.65.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Edelman G. M. Surface modulation in cell recognition and cell growth. Science. 1976 Apr 16;192(4236):218–226. doi: 10.1126/science.769162. [DOI] [PubMed] [Google Scholar]
  15. Fambrough D. M., Devreotes P. N. Newly synthesized acetylcholine receptors are located in the Golgi apparatus. J Cell Biol. 1978 Jan;76(1):237–244. doi: 10.1083/jcb.76.1.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fischbach G. D., Cohen S. A. The distribution of acetylcholine sensitivity over uninnervated and innervated muscle fibers grown in cell culture. Dev Biol. 1973 Mar;31(1):147–162. doi: 10.1016/0012-1606(73)90326-6. [DOI] [PubMed] [Google Scholar]
  17. Fischbach G. D. Synapse formation between dissociated nerve and muscle cells in low density cell cultures. Dev Biol. 1972 Jun;28(2):407–429. doi: 10.1016/0012-1606(72)90023-1. [DOI] [PubMed] [Google Scholar]
  18. Fischman D. A. The synthesis and assembly of myofibrils in embryonic muscle. Curr Top Dev Biol. 1970;5:235–280. doi: 10.1016/s0070-2153(08)60057-5. [DOI] [PubMed] [Google Scholar]
  19. Frank E., Fischbach G. D. Early events in neuromuscular junction formation in vitro: induction of acetylcholine receptor clusters in the postsynaptic membrane and morphology of newly formed synapses. J Cell Biol. 1979 Oct;83(1):143–158. doi: 10.1083/jcb.83.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Frank E., Gautvik K., Sommerschild H. Cholinergic receptors at denervated mammalian motor end-plates. Acta Physiol Scand. 1975 Sep;95(1):66–76. doi: 10.1111/j.1748-1716.1975.tb10026.x. [DOI] [PubMed] [Google Scholar]
  21. Hall Z. W., Lubit B. W., Schwartz J. H. Cytoplasmic actin in postsynaptic structures at the neuromuscular junction. J Cell Biol. 1981 Sep;90(3):789–792. doi: 10.1083/jcb.90.3.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hirokawa N., Heuser J. E. Internal and external differentiations of the postsynaptic membrane at the neuromuscular junction. J Neurocytol. 1982 Jun;11(3):487–510. doi: 10.1007/BF01257990. [DOI] [PubMed] [Google Scholar]
  23. Kelly A. M., Zacks S. I. The fine structure of motor endplate morphogenesis. J Cell Biol. 1969 Jul;42(1):154–169. doi: 10.1083/jcb.42.1.154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kuromi H., Kidokoro Y. Nerve disperses preexisting acetylcholine receptor clusters prior to induction of receptor accumulation in Xenopus muscle cultures. Dev Biol. 1984 May;103(1):53–61. doi: 10.1016/0012-1606(84)90006-x. [DOI] [PubMed] [Google Scholar]
  25. Lomo T., Rosenthal J. Control of ACh sensitivity by muscle activity in the rat. J Physiol. 1972 Mar;221(2):493–513. doi: 10.1113/jphysiol.1972.sp009764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lomo T., Westgaard R. H. Further studies on the control of ACh sensitivity by muscle activity in the rat. J Physiol. 1975 Nov;252(3):603–626. doi: 10.1113/jphysiol.1975.sp011161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lubit B. W. Association of beta-cytoplasmic actin with high concentrations of acetylcholine receptor (AChR) in normal and anti-AChR-treated primary rat muscle cultures. J Histochem Cytochem. 1984 Sep;32(9):973–981. doi: 10.1177/32.9.6379042. [DOI] [PubMed] [Google Scholar]
  28. McMahan U. J., Sanes J. R., Marshall L. M. Cholinesterase is associated with the basal lamina at the neuromuscular junction. Nature. 1978 Jan 12;271(5641):172–174. doi: 10.1038/271172a0. [DOI] [PubMed] [Google Scholar]
  29. Merlie J. P., Sanes J. R. Concentration of acetylcholine receptor mRNA in synaptic regions of adult muscle fibres. Nature. 1985 Sep 5;317(6032):66–68. doi: 10.1038/317066a0. [DOI] [PubMed] [Google Scholar]
  30. NOVIKOFF A. B., GOLDFISCHER S. Nucleosidediphosphatase activity in the Golgi apparatus and its usefulness for cytological studies. Proc Natl Acad Sci U S A. 1961 Jun 15;47:802–810. doi: 10.1073/pnas.47.6.802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nakai J. The development of neuromuscular junctions in cultures of chick embryo tissues. J Exp Zool. 1969 Jan;170(1):85–106. doi: 10.1002/jez.1401700108. [DOI] [PubMed] [Google Scholar]
  32. Peng H. B. Cytoskeletal organization of the presynaptic nerve terminal and the acetylcholine receptor cluster in cell cultures. J Cell Biol. 1983 Aug;97(2):489–498. doi: 10.1083/jcb.97.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Peng H. B., Froehner S. C. Association of the postsynaptic 43K protein with newly formed acetylcholine receptor clusters in cultured muscle cells. J Cell Biol. 1985 May;100(5):1698–1705. doi: 10.1083/jcb.100.5.1698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Peng H. B., Phelan K. A. Early cytoplasmic specialization at the presumptive acetylcholine receptor cluster: a meshwork of thin filaments. J Cell Biol. 1984 Jul;99(1 Pt 1):344–349. doi: 10.1083/jcb.99.1.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ravdin P., Axelrod D. Fluorescent tetramethyl rhodamine derivatives of alpha-bungarotoxin: preparation, separation, and characterization. Anal Biochem. 1977 Jun;80(2):585–592. doi: 10.1016/0003-2697(77)90682-0. [DOI] [PubMed] [Google Scholar]
  36. Role L. W., Matossian V. R., O'Brien R. J., Fischbach G. D. On the mechanism of acetylcholine receptor accumulation at newly formed synapses on chick myotubes. J Neurosci. 1985 Aug;5(8):2197–2204. doi: 10.1523/JNEUROSCI.05-08-02197.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rubin L. L. Increases in muscle Ca2+ mediate changes in acetylcholinesterase and acetylcholine receptors caused by muscle contraction. Proc Natl Acad Sci U S A. 1985 Oct;82(20):7121–7125. doi: 10.1073/pnas.82.20.7121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sanes J. R., Hall Z. W. Antibodies that bind specifically to synaptic sites on muscle fiber basal lamina. J Cell Biol. 1979 Nov;83(2 Pt 1):357–370. doi: 10.1083/jcb.83.2.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Walker J. H., Boustead C. M., Witzemann V. The 43-K protein, v1, associated with acetylcholine receptor containing membrane fragments is an actin-binding protein. EMBO J. 1984 Oct;3(10):2287–2290. doi: 10.1002/j.1460-2075.1984.tb02127.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ziskind-Conhaim L., Geffen I., Hall Z. W. Redistribution of acetylcholine receptors on developing rat myotubes. J Neurosci. 1984 Sep;4(9):2346–2349. doi: 10.1523/JNEUROSCI.04-09-02346.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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