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. 1988 Jun 1;106(6):2087–2093. doi: 10.1083/jcb.106.6.2087

Distribution of extrajunctional acetylcholine receptors on a vertebrate muscle: evaluated by using a scanning electron microscope autoradiographic procedure

PMCID: PMC2115121  PMID: 3384854

Abstract

A scanning electron microscope (SEM) autoradiographic technique was calibrated and used to determine the site density of acetylcholine receptors within 250 micron of the neuromuscular junction in innervated as well as 3- and 10-d denervated sternomastoid muscle of the mouse. In all these groups sharp gradients of receptor site density are seen around the endplates in the first 2-7 micron, continuing less sharply to between 25 and 50 micron. Beyond 50 micron (to 250 micron) a spatial density gradient is present 3 d after denervation, but none exist by 10 d. These results suggest that the postdenervation steady-state extrajunctional receptor site density is reached sooner near the junction than away from the junction. The usefulness of SEM autoradiography to study the expression and distribution of membrane molecules at high resolution is demonstrated.

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Selected References

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  1. AXELSSON J., THESLEFF S. A study of supersensitivity in denervated mammalian skeletal muscle. J Physiol. 1959 Jun 23;147(1):178–193. doi: 10.1113/jphysiol.1959.sp006233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Albuquerque E. X., McIsaac R. J. Fast and slow mammalian muscles after denervation. Exp Neurol. 1970 Jan;26(1):183–202. doi: 10.1016/0014-4886(70)90099-3. [DOI] [PubMed] [Google Scholar]
  3. BACHMANN L., SALPETER M. M. AUTORADIOGRAPHY WITH THE ELECTRON MICROSCOPE; A QUANTITATIVE EVALUATION. Lab Invest. 1965 Jun;14:1041–1053. [PubMed] [Google Scholar]
  4. Bachmann L., Salpeter M. M. Absolute sensitivity of electron microscope radioautography. J Cell Biol. 1967 May;33(2):299–305. doi: 10.1083/jcb.33.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bekoff A., Betz W. Properties of isolated adult rat muscle fibres maintained in tissue culture. J Physiol. 1977 Oct;271(2):537–547. doi: 10.1113/jphysiol.1977.sp012013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. CARO L. G., VAN TUBERGEN R. P., KOLB J. A. High-resolution autoradiography. I. Methods. J Cell Biol. 1962 Nov;15:173–188. doi: 10.1083/jcb.15.2.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chang C. C., Chuang S. T., Huang M. C. Effects of chronic treatment with various neuromuscular blocking agents on the number and distribution of acetylcholine receptors in the rat diaphragm. J Physiol. 1975 Aug;250(1):161–173. doi: 10.1113/jphysiol.1975.sp011047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Desaki J., Uehara Y. The overall morphology of neuromuscular junctions as revealed by scanning electron microscopy. J Neurocytol. 1981 Feb;10(1):101–110. doi: 10.1007/BF01181747. [DOI] [PubMed] [Google Scholar]
  9. Fambrough D. M. Acetylcholine receptors. Revised estimates of extrajunctional receptor density in denervated rat diaphragm. J Gen Physiol. 1974 Oct;64(4):468–472. doi: 10.1085/jgp.64.4.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fambrough D. M. Control of acetylcholine receptors in skeletal muscle. Physiol Rev. 1979 Jan;59(1):165–227. doi: 10.1152/physrev.1979.59.1.165. [DOI] [PubMed] [Google Scholar]
  11. Fertuck H. C., Salpeter M. M. Quantitation of junctional and extrajunctional acetylcholine receptors by electron microscope autoradiography after 125I-alpha-bungarotoxin binding at mouse neuromuscular junctions. J Cell Biol. 1976 Apr;69(1):144–158. doi: 10.1083/jcb.69.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fertuck H. C., Salpeter M. M. Sensitivity in electron microscope autoradiography for 125I. J Histochem Cytochem. 1974 Feb;22(2):80–87. doi: 10.1177/22.2.80. [DOI] [PubMed] [Google Scholar]
  13. Fertuck H. C., Woodward W., Salpeter M. M. In vivo recovery of muscle contraction after alpha-bungarotoxin binding. J Cell Biol. 1975 Jul;66(1):209–213. doi: 10.1083/jcb.66.1.209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harris W. V., Salpeter M. M. Soluble compound electron microscope (EM) autoradiography: a resolution source to test redistribution of soluble tritiated compounds during processing. J Histochem Cytochem. 1983 Apr;31(4):495–500. doi: 10.1177/31.4.6827081. [DOI] [PubMed] [Google Scholar]
  15. Hartzell H. C., Fambrough D. M. Acetylcholine receptors. Distribution and extrajunctional density in rat diaphragm after denervation correlated with acetylcholine sensitivity. J Gen Physiol. 1972 Sep;60(3):248–262. doi: 10.1085/jgp.60.3.248. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Junger E., Bachmann L. Methodological basis for an autoradiographic demonstration of insulin receptor sites on the surface of whole cells: a study using light- and scanning electron microscopy. J Microsc. 1980 Jul;119(2):199–211. doi: 10.1111/j.1365-2818.1980.tb04090.x. [DOI] [PubMed] [Google Scholar]
  17. Kuffler S. W., Yoshikami D. The distribution of acetylcholine sensitivity at the post-synaptic membrane of vertebrate skeletal twitch muscles: iontophoretic mapping in the micron range. J Physiol. 1975 Jan;244(3):703–730. doi: 10.1113/jphysiol.1975.sp010821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Land B. R., Podleski T. R., Salpeter E. E., Salpeter M. M. Acetylcholine receptor distribution on myotubes in culture correlated to acetylcholine sensitivity. J Physiol. 1977 Jul;269(1):155–176. doi: 10.1113/jphysiol.1977.sp011897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lane M. A., Sastre A., Law M., Salpeter M. M. Cholinergic and adrenergic receptors on mouse cardiocytes in vitro. Dev Biol. 1977 Jun;57(2):254–269. doi: 10.1016/0012-1606(77)90213-5. [DOI] [PubMed] [Google Scholar]
  20. Levitt-Gilmour T. A., Salpeter M. M. Gradient of extrajunctional acetylcholine receptors early after denervation of mammalian muscle. J Neurosci. 1986 Jun;6(6):1606–1612. doi: 10.1523/JNEUROSCI.06-06-01606.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Loring R. H., Salpeter M. M. Denervation increases turnover rate of junctional acetylcholine receptors. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2293–2297. doi: 10.1073/pnas.77.4.2293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. MILEDI R. The acetylcholine sensitivity of frog muscle fibres after complete or partial devervation. J Physiol. 1960 Apr;151:1–23. [PMC free article] [PubMed] [Google Scholar]
  23. Matthews-Bellinger J. A., Salpeter M. M. Fine structural distribution of acetylcholine receptors at developing mouse neuromuscular junctions. J Neurosci. 1983 Mar;3(3):644–657. doi: 10.1523/JNEUROSCI.03-03-00644.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Matthews-Bellinger J., Salpeter M. M. Distribution of acetylcholine receptors at frog neuromuscular junctions with a discussion of some physiological implications. J Physiol. 1978 Jun;279:197–213. doi: 10.1113/jphysiol.1978.sp012340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Neugebauer K., Salpeter M. M., Podleski T. R. Differential responses of L5 and rat primary muscle cells to factors in rat brain extract. Brain Res. 1985 Oct 28;346(1):58–69. doi: 10.1016/0006-8993(85)91094-7. [DOI] [PubMed] [Google Scholar]
  26. Paul D., Gröbe A., Zimmer F. Autoradiography in the scanning electron microscope. Nature. 1970 Aug 1;227(5257):488–489. doi: 10.1038/227488a0. [DOI] [PubMed] [Google Scholar]
  27. Pestronk A., Drachman D. B., Griffin J. W. Effect of muscle disuse on acetylcholine receptors. Nature. 1976 Mar 25;260(5549):352–353. doi: 10.1038/260352a0. [DOI] [PubMed] [Google Scholar]
  28. SALPETER M. M., BACHMANN L. AUTORADIOGRAPHY WITH THE ELECTRON MICROSCOPE. A PROCEDURE FOR IMPROVING RESOLUTION, SENSITIVITY, AND CONTRAST. J Cell Biol. 1964 Aug;22:469–477. doi: 10.1083/jcb.22.2.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Salpeter M. M., Budd G. C., Mattimoe S. Resolution in autoradiography using semithin sections. J Histochem Cytochem. 1974 Apr;22(4):217–222. doi: 10.1177/22.4.217. [DOI] [PubMed] [Google Scholar]
  30. Salpeter M. M., Fertuck H. C., Salpeter E. E. Resolution in electron microscope autoradiography. III. Iodine-125, the effect of heavy metal staining, and a reassessment of critical parameters. J Cell Biol. 1977 Jan;72(1):161–173. doi: 10.1083/jcb.72.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Salpeter M. M., Nelson R., Harris R., Salpeter E. E. Resolution in EM autoradiography V: 45Ca (and 33P). J Histochem Cytochem. 1987 Oct;35(10):1047–1051. doi: 10.1177/35.10.3624849. [DOI] [PubMed] [Google Scholar]
  32. Salpeter M. M. Sensitivity in electron microscope autoradiography. II. Effect of heavy metal staining. J Histochem Cytochem. 1973 Jul;21(7):623–627. doi: 10.1177/21.7.623. [DOI] [PubMed] [Google Scholar]
  33. Salpeter M. M., Szabo M. Sensitivity in electron microscope autoradiography. I. The effect of radiation dose. J Histochem Cytochem. 1972 Jun;20(6):425–434. doi: 10.1177/20.6.425. [DOI] [PubMed] [Google Scholar]
  34. Steinbach J. H. Neuromuscular junctions and alpha-bungarotoxin-binding sites in denervated and contralateral cat skeletal muscles. J Physiol. 1981;313:513–528. doi: 10.1113/jphysiol.1981.sp013679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Weiss R. L. Scanning electron microscope autoradiography of critical point dried biological samples. Scan Electron Microsc. 1980;(4):123–130. [PubMed] [Google Scholar]

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