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. 1971 Dec 1;51(3):703–721. doi: 10.1083/jcb.51.3.703

THE APPEARANCE OF ACETYLCHOLINESTERASE IN THE MYOTOME OF THE EMBRYONIC RABBIT

An Electron Microscope Cytochemical and Biochemical Study

Virginia M Tennyson 1, Miro Brzin 1, Paul Slotwiner 1
PMCID: PMC2108035  PMID: 4256859

Abstract

Acetylcholinesterase (AChE) activity has been studied in the myoblast of skeletal muscle of the 9–13 day fetal rabbit. Cytochemical activity is present in the nuclear envelope and the endoplasmic reticulum, including its derivatives the subsurface reticulum and the sarcoplasmic reticulum. End product is also found in the Golgi complex of the more differentiated myoblasts. The formation of reticulum-bound acetylcholinesterase in the myoblast appears to be independent of nerve-muscle contact, since the enzyme is present before the outgrowth of the spinal nerve. The nerve lacks cytochemical end product until the myoblast is well differentiated. Possible mechanisms of spontaneous muscle contraction have been discussed. A second type of myotomal cell, which exhibits a poorly localized end product of AChE activity, has been described. The ready solubility of the enzyme or diffusibility of its end product suggests that the enzyme may be a lyoesterase. This cell may be the precursor of the morphologically undifferentiated cell which is closely apposed to the myotubes in later stages of skeletal muscle development. Biochemical studies show a significant increase in AChE activity in the dermomyotome by day 12, when many of the myoblasts are well differentiated and the second type of myotomal cell is prominent. Cytochemical studies have indicated that many of the cells in the sample lack reaction product of enzymic activity, whereas others are very active. Biochemical values, therefore, reflect the amount of enzyme in the dermomyotome as a whole, but give little information on the enzymic content of individual cells.

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Selected References

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  1. ALLEN E. R., PEPE F. A. ULTRASTRUCTURE OF DEVELOPING MUSCLE CELLS IN THE CHICK EMBRYO. Am J Anat. 1965 Jan;116:115–147. doi: 10.1002/aja.1001160107. [DOI] [PubMed] [Google Scholar]
  2. BERGMAN R. A. Observations on the morphogenesis of rat skeletal muscle. Bull Johns Hopkins Hosp. 1962 Apr;110:187–201. [PubMed] [Google Scholar]
  3. BERNSOHN J., BARRON K. D., HESS A. R. Multiple nature of acetylcholinesterase in nerve tissue. Nature. 1962 Jul 21;195:285–286. doi: 10.1038/195285a0. [DOI] [PubMed] [Google Scholar]
  4. Bellhorn M. B., Blumenfeld O. O., Gallop P. M. Acetylcholinesterase of the human erythrocyte membrane. Biochem Biophys Res Commun. 1970 Apr 24;39(2):267–273. doi: 10.1016/0006-291x(70)90788-6. [DOI] [PubMed] [Google Scholar]
  5. Burt A. M. Acetylcholinesterase and choline acetyltransferase activity in the developing chick spinal cord. J Exp Zool. 1968 Sep;169(1):107–112. doi: 10.1002/jez.1401690112. [DOI] [PubMed] [Google Scholar]
  6. Church J. C. Satellite cells and myogenesis; a study in the fruit-bat web. J Anat. 1969 Nov;105(Pt 3):419–438. [PMC free article] [PubMed] [Google Scholar]
  7. DESSOUKY D. A., HIBBS R. G. AN ELECTRON MICROSCOPE STUDY OF THE DEVELOPMENT OF THE SOMATIC MUSCLE OF THE CHICK EMBRYO. Am J Anat. 1965 May;116:523–565. doi: 10.1002/aja.1001160305. [DOI] [PubMed] [Google Scholar]
  8. ERAENKOE O., HAERKOENEN M., KOKKO A., RAEISAENEN L. HISTOCHEMICAL AND STARCH GEL ELECTROPHORETIC CHARACTERIZATION OF DESMO- AND LYO-ESTERASES IN THE SYMPATHETIC AND SPINAL GANGLIA OF THE RAT. J Histochem Cytochem. 1964 Aug;12:570–581. doi: 10.1177/12.8.570. [DOI] [PubMed] [Google Scholar]
  9. Ecobichon D. J., Israel Y. Characterization of the esterases from electric tissue of Electrophorus by starch-gel electrophoresis. Can J Biochem. 1967 Jul;45(7):1099–1105. doi: 10.1139/o67-127. [DOI] [PubMed] [Google Scholar]
  10. Fischman D. A. An electron microscope study of myofibril formation in embryonic chick skeletal muscle. J Cell Biol. 1967 Mar;32(3):557–575. doi: 10.1083/jcb.32.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. GEREBTZOFF M. A. L'appareil cholinestérasique musculo-tendineux; structure, développment, effet de la dénervation et de la ténotomie. Acta Physiol Pharmacol Neerl. 1957;6:419–427. [PubMed] [Google Scholar]
  12. HAMBURGER V. SOME ASPECTS OF THE EMBRYOLOGY OF BEHAVIOR. Q Rev Biol. 1963 Dec;38:342–365. doi: 10.1086/403941. [DOI] [PubMed] [Google Scholar]
  13. HAY E. D. The fine structure of differentiating muscle in the salamander tail. Z Zellforsch Mikrosk Anat. 1963;59:6–34. doi: 10.1007/BF00321005. [DOI] [PubMed] [Google Scholar]
  14. Hagopian M., Tennyson V. M., Spiro D. Cytochemical localization of cholinesterase in embryonic rabbit cardiac muscle. J Histochem Cytochem. 1970 Jan;18(1):38–43. doi: 10.1177/18.1.38. [DOI] [PubMed] [Google Scholar]
  15. Hamburger V., Balaban M., Oppenheim R., Wenger E. Periodic motility of normal and spinal chick embryos between 8 and 17 days of incubation. J Exp Zool. 1965 Jun;159(1):1–13. doi: 10.1002/jez.1401590102. [DOI] [PubMed] [Google Scholar]
  16. Hirano H. Ultrastructural study on the morphogenesis of the neuromuscular junction in the skeletal muscle of the chick. Z Zellforsch Mikrosk Anat. 1967;79(2):198–208. [PubMed] [Google Scholar]
  17. Ishikawa H., Bischoff R., Holtzer H. Mitosis and intermediate-sized filaments in developing skeletal muscle. J Cell Biol. 1968 Sep;38(3):538–555. doi: 10.1083/jcb.38.3.538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ishikawa H. Electron microscopic observations of satellite cells with special reference to the development of mammalian skeletal muscles. Z Anat Entwicklungsgesch. 1966;125(1):43–63. doi: 10.1007/BF00521974. [DOI] [PubMed] [Google Scholar]
  19. Ishikawa H. Formation of elaborate networks of T-system tubules in cultured skeletal muscle with special reference to the T-system formation. J Cell Biol. 1968 Jul;38(1):51–66. doi: 10.1083/jcb.38.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. KOELLE G. B., FRIEDENWALD J. A. A histochemical method for localizing cholinesterase activity. Proc Soc Exp Biol Med. 1949 Apr;70(4):617–622. doi: 10.3181/00379727-70-17013. [DOI] [PubMed] [Google Scholar]
  21. KOELLE G. B. The elimination of enzymatic diffusion artifacts in the histochemical localization of cholinesterases and a survey of their cellular distributions. J Pharmacol Exp Ther. 1951 Oct;103(2):153–171. [PubMed] [Google Scholar]
  22. KUPFER C., KOELLE G. B. A histochemical study of cholinesterase during formation of the motor end plate of the albino rat. J Exp Zool. 1951 Apr;116(3):397–413. doi: 10.1002/jez.1401160303. [DOI] [PubMed] [Google Scholar]
  23. Kelly A. M., Zacks S. I. The histogenesis of rat intercostal muscle. J Cell Biol. 1969 Jul;42(1):135–153. doi: 10.1083/jcb.42.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Koelle W. A., Hossaini K. S., Akbarzadeh P., Koelle G. B. Histochemical evidence and consequences of the occurrence of isoenzymes of acetylcholinesterase. J Histochem Cytochem. 1970 Nov;18(11):812–819. doi: 10.1177/18.11.812. [DOI] [PubMed] [Google Scholar]
  25. LI C. L., ENGEL K., KLATZO I. Some properties of cultured chick skeletal muscle with particular reference to fibrillation potential. J Cell Comp Physiol. 1959 Jun;53:421–444. doi: 10.1002/jcp.1030530307. [DOI] [PubMed] [Google Scholar]
  26. Langley O. K., Landon D. N. Copper binding at nodes of Ranvier: a new electron histochemical technique for the demonstration of polyanions. J Histochem Cytochem. 1969 Jan;17(1):66–69. doi: 10.1177/17.1.66. [DOI] [PubMed] [Google Scholar]
  27. Lentz T. L. Development of the neuromuscular junction. I. Cytological and cytochemical studies on the neuromuscular junction of differentiating muscle in the regenerating limb of the newt Triturus. J Cell Biol. 1969 Aug;42(2):431–443. doi: 10.1083/jcb.42.2.431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. MAURO A. Satellite cell of skeletal muscle fibers. J Biophys Biochem Cytol. 1961 Feb;9:493–495. doi: 10.1083/jcb.9.2.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. MUMENTHALER M., ENGEL W. K. Cytological localization of cholinesterase in developing chick embryo skeletal muscle. Acta Anat (Basel) 1961;47:274–299. doi: 10.1159/000141814. [DOI] [PubMed] [Google Scholar]
  30. McCaman M. W., Tomey L. R., McCaman R. E. Radiomimetric assay of acetylcholinesterase activity in submicrogram amounts of tissue. Life Sci. 1968 Mar 15;7(6):233–244. doi: 10.1016/0024-3205(68)90017-9. [DOI] [PubMed] [Google Scholar]
  31. Muir A. R., Kanji A. H., Allbrook D. The structure of the satellite cells in skeletal muscle. J Anat. 1965 Jul;99(Pt 3):435–444. [PMC free article] [PubMed] [Google Scholar]
  32. Nachmansohn D. Proteins in excitable membranes: their properties and function in bioelectricity are discussed. Science. 1970 May 29;168(3935):1059–1066. doi: 10.1126/science.168.3935.1059. [DOI] [PubMed] [Google Scholar]
  33. PALADE G. E. A study of fixation for electron microscopy. J Exp Med. 1952 Mar;95(3):285–298. doi: 10.1084/jem.95.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Przybylski R. J., Blumberg J. M. Ultrastructural aspects of myogenesis in the chick. Lab Invest. 1966 May;15(5):836–863. [PubMed] [Google Scholar]
  35. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. ROSENBLUTH J. Subsurface cisterns and their relationship to the neuronal plasma membrane. J Cell Biol. 1962 Jun;13:405–421. doi: 10.1083/jcb.13.3.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Richter D., Croft P. G. Blood esterases. Biochem J. 1942 Dec;36(10-12):746–757. doi: 10.1042/bj0360746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Robbins N., Yonezawa T. Developing neuromuscular junctions: first signs of chemical transmission during formation in tissue culture. Science. 1971 Apr 23;172(3981):395–398. doi: 10.1126/science.172.3981.395. [DOI] [PubMed] [Google Scholar]
  39. Romanes G. J. The development and significance of the cell columns in the ventral horn of the cervical and upper thoracic spinal cord of the rabbit. J Anat. 1941 Oct;76(Pt 1):112–130.5. [PMC free article] [PubMed] [Google Scholar]
  40. SABATINI D. D., BENSCH K., BARRNETT R. J. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol. 1963 Apr;17:19–58. doi: 10.1083/jcb.17.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tennyson V. M., Brzin M. The appearance of acetylcholinesterase in the dorsal root neuroblast of the rabbit embryo. A study by electron microscope cytochemistry and microgasometric analysis with the magnetic diver. J Cell Biol. 1970 Jul;46(1):64–80. doi: 10.1083/jcb.46.1.64. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Tennyson V. M. The fine structure of the axon and growth cone of the dorsal root neuroblast of the rabbit embryo. J Cell Biol. 1970 Jan;44(1):62–79. doi: 10.1083/jcb.44.1.62. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Veneroni G., Murray M. R. Formation de novo and development of neuromuscular junctions in vitro. J Embryol Exp Morphol. 1969 Apr;21(2):369–382. [PubMed] [Google Scholar]
  44. WATSON M. L. Staining of tissue sections for electron microscopy with heavy metals. J Biophys Biochem Cytol. 1958 Jul 25;4(4):475–478. doi: 10.1083/jcb.4.4.475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. ZACKS S. I. Esterases in the early chick embryo. Anat Rec. 1954 Mar;118(3):509–537. doi: 10.1002/ar.1091180304. [DOI] [PubMed] [Google Scholar]

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