Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Localization of acetylcholinesterase in the rat myoneural junction

  • 18 Accesses

  • 17 Citations

Summary

The distribution of myoneural acetylcholinesterase (AChE; EC. 3.1.1.7) was studied electron microscopically with the copper ferricyanide method at pH 6.0, using acetylthiocholine iodide as substrate and iso-OMPA to exclude other cholinesterase activity.

It was observed that the results obtained with this method are affected by changes in the reaction temperature, inhibitor concentration and fixation time. Changes which retard the rate of hydrolysis of AChE were observed to transfer the localization of the reaction endproduct in intact junctions from the extracellular side of the postsynaptic membrane to the intracellular side of the postsynaptic membrane. From the results it is concluded that the site of most intense AChE corresponds to the region of increased subsarcolemmal electron density of the postsynaptic membrane.

This is a preview of subscription content, log in to check access.

References

  1. Barrnett, R. J.: The fine structural localization of acetylcholinesterase at the myoneural junction. J. Cell Biol. 12, 247–262 (1962).

  2. Bell, C.: Use of the direct-coloring thiocholine technique for demonstration of intracellular neuronal cholinesterases. J. Histochem. Cytochem. 14, 567–570 (1966).

  3. Bergman, R. A., H. Ueno, Y. Morizono, J. S. Hanker, and A. M. Seligman: Ultrastructural demonstration of acetylcholinesterase activity of motor end plates via osmiophilic diazothioethers. Histochemie 11, 1–12 (1967).

  4. Brown, L. M.: A thiolcholine method for locating cholinesterase activity by electron microscopy. Bibl. anat. (Basel) 2, 21–33 (1961).

  5. Couteaux, R.: Morphological and cytochemical observations on the postsynaptic membrane at motor end plates and ganglionic synapses. Exp. Cell Res., Suppl. 5, 294–322 (1958).

  6. Davis, R., and G. B. Koelle: Electron microscopic localization of acetylcholinesterase at the motor end plate by the Gold-Thiolacetic acid and Gold-Thiocholine methods. J. Histochem. Cytochem. 13, 703–704 (1965).

  7. —: Electron microscopic localization of acetylcholinesterase and non-specific cholinesterase at the neuromuscular junction by the gold-thiocholine and gold-thiolacetic acid methods. J. Cell Biol. 34, 157–171 (1967).

  8. De Robertis, E., A. Pellegrino de Iraldi, G. Rodriquez de Lores Arnaiz, and L. Salganicoff: Cholinergic and non-cholinergic nerve endings in rat brain. I. Isolation and subcellular distribution of acetylcholine and acetylcholinesterase. J. Neurochem. 9, 23–35 (1962).

  9. Eränkö, O., L. Rechardt, and L. Hänninen: Electron microscopic demonstration of cholinesterases in nervous tissue. Histochemie 8, 369–376 (1967).

  10. Goedde, H. W., A. Doenicke, and K. Altland (eds.): In: Pseudocholinesterases. Berlin-Heidelberg-New York: Springer 1967.

  11. Hirano, H.: A histochemical study of the cholinesterase activity in the neuromuscular junction in developing chick skeletal muscles. Arch. histol. jap. 28, 89–101 (1967).

  12. Iwayama, T.: Electron microscopic study on cholinesterase at the neuromuscular junction. Okajimas Folia anat. jap. 42, 329–359 (1966).

  13. Karczmar, A. G.: Neuromuscular pharmacology. Ann. Rev. Pharmacol. 7, 231–276 (1967).

  14. Karlin, A.: The association of acetylcholinesterase and membrane in subcellular fractions of the electric tissue of Electrophorus. J. Cell Biol. 25, 159–169 (1965).

  15. Karnovsky, M. J.: The localization of cholinesterase activity in rat cardiac muscle by electron microscopy. J. Cell Biol. 23, 217–232 (1964).

  16. —, and L. Roots: A “direct colouring” thiocholine method for cholinesterases. J. Histochem. Cytochem. 12, 219–221 (1964).

  17. Kása, P., and B. Csillik: Electron microscopic localization of cholinesterase by a copperlead-thiocholine technique. J. Neurochem. 13, 1345–1349 (1966).

  18. —: AChE synthesis in cholinergic neurons: Electron histochemistry of enzyme translocation. Histochemie 12, 175–183 (1968).

  19. Lehrer, G. M., and L. Ornstein: A diazo coupling method for the electron microscopic localization of cholinesterase. J. biophys. biochem. Cytol. 6, 399–406 (1959).

  20. Lewis, P. R., and C. C. D. Shute: Demonstration of cholinesterase activity with the electron microscope. J. Physiol. (Lond.) 175, 5P-7P (1964).

  21. —: The distribution of cholinesterase in cholinergic neurons demonstrated with the electron microscopy. J. Cell Sci. 1, 381–390 (1966).

  22. Luft, J. H.: Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9, 409–414 (1961).

  23. Miledi, R.: Electron microscopical localization of products from histochemical reactions used to detect cholinesterase in muscle. Nature (Lond.) 204, 293–295 (1964).

  24. Reynolds, E. S.: The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J. Cell Biol. 17, 208–212 (1963).

  25. Sabatini, D. D., K. Bensch, and R. J. Barrnett: Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J. Cell Biol. 17, 19–58 (1963).

  26. Salpeter, M. M.: Electron microscope radioautography as a quantitative tool in enzyme cytochemistry. J. Cell Biol. 32, 379–389 (1967).

  27. Schlaepfer, W. W., and R. M. Torack: The ultrastructural localization of cholinesterase activity in the sciatic nerve of the rat. J. Histochem. Cytochem. 14, 369–378 (1966).

  28. Teräväinen, H.: Electron microscopic localization of cholinesterases in the rat myoneural junction. Histochemie 10, 266–271 (1967).

  29. —: Light and electron microscopic observations on the distribution of carboxylic esterases in adult, developing, degenerating and regenerating myoneural junctions of the rat. Academic Diss. Painovalmiste, Helsinki (1968).

  30. Toschi, G.: A biochemical study of brain microsomes. Exp. Cell Res. 16, 232–255 (1959).

  31. Whittaker, V. P., J. A. Michaelson, and R. J. A. Kirkland: The separation of synaptic vesicles from nerve-ending particles (“synaptosomes”). Biochem. J. 90, 293–303 (1964).

  32. Zacks, S. I., and J. M. Blumberg: The histochemical localization of acetylcholinesterase in the fine structure of neuromuscular junctions of mouse and human intestinal muscle. J. Histochem. Cytochem. 9, 317–324 (1961).

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Teräväinen, H. Localization of acetylcholinesterase in the rat myoneural junction. Histochemie 17, 162–169 (1969). https://doi.org/10.1007/BF00277782

Download citation

Keywords

  • Public Health
  • Copper
  • Iodide
  • Reaction Temperature
  • Inhibitor Concentration