Localization of actin, beta-spectrin, 43 x 10(3) Mr and 58 x 10(3) Mr proteins to receptor-enriched domains of newly formed acetylcholine receptor aggregates in isolated myotube membranes - PubMed (original) (raw)
. 1990 Dec:97 ( Pt 4):615-26.
doi: 10.1242/jcs.97.4.615.
Affiliations
- PMID: 2127596
- DOI: 10.1242/jcs.97.4.615
Localization of actin, beta-spectrin, 43 x 10(3) Mr and 58 x 10(3) Mr proteins to receptor-enriched domains of newly formed acetylcholine receptor aggregates in isolated myotube membranes
M P Daniels. J Cell Sci. 1990 Dec.
Abstract
I have examined the possible involvement of specific cytoskeletal and peripheral membrane proteins in the early stages of acetylcholine receptor (AChR) aggregation in rat myotubes in culture by immunofluorescence localization of these proteins on the cytoplasmic face of isolated plasma membranes. A culture procedure utilizing selective replating of myoblasts and subsequent treatment with cytosine arabinoside was devised to obtain large, multipolar myotubes with extensive upper surfaces that are free of fibroblasts. These cultures were exposed for 4-6 h to embryonic pig brain extract (EBX) to induce AChR aggregate formation on the upper cell surface, and the AChRs were labeled with TRITC-conjugated alpha-bungarotoxin. Large sheets of plasma membranes from the upper cell surface were isolated by adhesion to a coverslip coated with a polypeptide adhesive (Cell-Tak) that was pressed on top of the culture. The membranes were labeled by indirect immunofluorescence with monoclonal antibodies against the 43 x 10(3) Mr and 58 x 10(3) Mr proteins, originally identified in the AChR-enriched membranes of Torpedo electroplaques, and with monoclonal antibodies against isoforms of actin and beta-spectrin. The labeling patterns showed that all four of these proteins are concentrated in the punctate AChR-enriched domains within the aggregates, suggesting that they may be involved in the early stages of AChR aggregation. Immunofluorescence labeling with monoclonal antibodies against vinculin and clathrin, and with an antiserum to talin, showed that these proteins are also associated with AChR aggregates; however, their labeling patterns did not correspond closely to the AChR-enriched domains. Furthermore, vinculin and talin dissociated from most of the membrane during isolation. The concentration of beta-spectrin and actin isoforms on the cytoplasmic fact of the AChR-enriched domains is consistent with the formation, early in the aggregation process, of a membrane-cytoskeleton association similar to that of erythrocytes.
Similar articles
- Association of cytoskeletal proteins with newly formed acetylcholine receptor aggregates induced by embryonic brain extract.
Daniels MP, Krikorian JG, Olek AJ, Bloch RJ. Daniels MP, et al. Exp Cell Res. 1990 Jan;186(1):99-108. doi: 10.1016/0014-4827(90)90215-v. Exp Cell Res. 1990. PMID: 2105221 - Acetylcholine receptor clusters of rat myotubes have at least three domains with distinctive cytoskeletal and membranous components.
Pumplin DW. Pumplin DW. J Cell Biol. 1989 Aug;109(2):739-53. doi: 10.1083/jcb.109.2.739. J Cell Biol. 1989. PMID: 2760110 Free PMC article. - An unusual beta-spectrin associated with clustered acetylcholine receptors.
Bloch RJ, Morrow JS. Bloch RJ, et al. J Cell Biol. 1989 Feb;108(2):481-93. doi: 10.1083/jcb.108.2.481. J Cell Biol. 1989. PMID: 2645300 Free PMC article. - Actin-membrane interaction in fibroblasts: what proteins are involved in this association?
Mangeat P, Burridge K. Mangeat P, et al. J Cell Biol. 1984 Jul;99(1 Pt 2):95s-103s. doi: 10.1083/jcb.99.1.95s. J Cell Biol. 1984. PMID: 6430913 Free PMC article. Review. - Recombinant neuromuscular synapses.
Phillips WD, Merlie JP. Phillips WD, et al. Bioessays. 1992 Oct;14(10):671-9. doi: 10.1002/bies.950141005. Bioessays. 1992. PMID: 1365879 Review.
Cited by
- Tissue engineering the mechanosensory circuit of the stretch reflex arc: sensory neuron innervation of intrafusal muscle fibers.
Rumsey JW, Das M, Bhalkikar A, Stancescu M, Hickman JJ. Rumsey JW, et al. Biomaterials. 2010 Nov;31(32):8218-27. doi: 10.1016/j.biomaterials.2010.07.027. Epub 2010 Aug 13. Biomaterials. 2010. PMID: 20708792 Free PMC article. - Tissue engineering intrafusal fibers: dose- and time-dependent differentiation of nuclear bag fibers in a defined in vitro system using neuregulin 1-beta-1.
Rumsey JW, Das M, Kang JF, Wagner R, Molnar P, Hickman JJ. Rumsey JW, et al. Biomaterials. 2008 Mar;29(8):994-1004. doi: 10.1016/j.biomaterials.2007.10.042. Biomaterials. 2008. PMID: 18076984 Free PMC article. - Calcium-activated proteases are critical for refilling depleted vesicle stores in cultured sensory-motor synapses of Aplysia.
Khoutorsky A, Spira ME. Khoutorsky A, et al. Learn Mem. 2005 Jul-Aug;12(4):414-22. doi: 10.1101/lm.92105. Learn Mem. 2005. PMID: 16077020 Free PMC article. - Effects of electrostimulation on glycogenolysis in cultured rat myotubes.
Elsner P, Grunnet N, Quistorff B. Elsner P, et al. Pflugers Arch. 2003 Dec;447(3):356-62. doi: 10.1007/s00424-003-1160-6. Epub 2003 Oct 25. Pflugers Arch. 2003. PMID: 14579114 - Synapse-forming axons and recombinant agrin induce microprocess formation on myotubes.
Uhm CS, Neuhuber B, Lowe B, Crocker V, Daniels MP. Uhm CS, et al. J Neurosci. 2001 Dec 15;21(24):9678-89. doi: 10.1523/JNEUROSCI.21-24-09678.2001. J Neurosci. 2001. PMID: 11739577 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous