Skip to main content
Log in

A histochemical method for the demonstration of acetylcholinesterase activity using semipermeable membranes

  • Original Articles
  • Published:
Histochemistry Aims and scope Submit manuscript

Summary

The “direct coloring” thiocholine method of Karnovsky and Roots (1964) for the demonstration of acetylcholinesterase (AChE) activity was modified and adapted to the technique of semipermeable membranes. In this way it is possible to demonstrate histochemically both the bound as well as the soluble part of AChE activity. The localization of the reaction product is very distinct. Microdensitometric investigations of results of this method showed a linear increase of the amount of reaction product up to an incubation time of 180 min and section thickness up to 24 μm. The medium supplemented with buffer (instead of agar) can be used for the demonstration of AChE activity in cryostat sections adherent to slides and is also very suitable for the defection of multiple forms of AChE in polyacrylamide or agarose gels.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Andrä J, Duijn P van (1985) Quantitative aspects of cytochemical methods for acetylcholinesterase studied with a cytochemical model system. Histochemistry 83:443–449

    Google Scholar 

  • Andrä J, Lojda Z, Weiss J, Luppa H (1985) Histochemical demonstration of soluble and fixation-labile acetylcholinesterase activity in the optic tectum of rudd and frog with a semipermeable membrane technique. Histochem J 17:572–575

    Google Scholar 

  • Bajgar J, Patočka J, Ornst F, Žižkovský V, Lacina P (1971) Subcellular distribution of acetylcholinesterase activity in human brain caudate nucleus. J Neurochem 18:529–530

    Google Scholar 

  • Eränkö O, Härkönen M, Kokko A, Räisänen L (1964) Histochemical and starch gel electrophoretic characterization of desmo- and lyoesterases in the sympathetic and spinal ganglia of the rat. J Histochem Cytochem 12:570–581

    Google Scholar 

  • Giacobini E (1960) The intracellular localization of cholinesterases. J Histochem Cytochem 8:419–424

    Google Scholar 

  • Hollunger EG, Niklasson BH (1973) The release and molecular state of mammalian brain acetylcholinesterase. J Neurochem 20:821–836

    Google Scholar 

  • Hüther G, Luppa H (1977) Demonstration of acetylcholinesterase by semipermeable membrane technique: Estimation of soluble and fixation-labile portions in different regions of the central nervous system. Histochemistry 51:245–251

    Google Scholar 

  • Hüther G, Luppa H (1979) The multiple forms of brain acetylcholinesterase III. Implications for the histochemical demonstration of acetylcholinesterase. Histochemistry 63:115–121

    Google Scholar 

  • Karnovsky MJ, Roots L (1964) A “direct coloring” thiocholine method for cholinesterase. J Histochem Cytochem 12:219–221

    Google Scholar 

  • Koelle GB, Friedenwald JS (1949) A histochemical method for localizing cholinesterase activity. Proc Soc Exp Biol (NY) 70:617–622

    Google Scholar 

  • Koelle WA, Hossaini KS, Akbarzadeh P, Koelle GB (1970) Histochemical evidence and consequences of the occurrence of isoenzymes of acetylcholinesterase. J Histochem Cytochem 18:812–819

    Google Scholar 

  • Lawler H (1964) The preparation of a soluble acetylcholinesterase from brain. Biochim Biophys Acta 81:280–288

    Google Scholar 

  • Lewis PR (1961) The effect of varying the conditions in the Koelle technique. Bibl Anat 2:11–20

    Google Scholar 

  • Lewis PR, Knight DP (1977) Staining methods for sectioned material. North Holland, Amsterdam

    Google Scholar 

  • Lojda Z, Gossrau R, Schiebler TH (1979) Enzyme histochemistry. A laboratory manual. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Lukáš Z, Fialová Z (1976) The possibilities and limitation of membrane methods for the histochemical demonstration of cholinesterases. Histochemistry 46:245–248

    Google Scholar 

  • Massoulié J, Bon S (1982) The molecular forms of cholinesterase and acetylcholinesterase in vertebrates. Annu Rev Neurosci 5:57–106

    Google Scholar 

  • Meijer AEFH (1980) Semipermeable membrane techniques in quantitative enzyme histochemistry. In: Trends in enzyme histochemistry and cytochemistry. Ciba Foundation Symposium 73 (New Series). Excerpta Medica, Amsterdam Oxford New York

    Google Scholar 

  • Nathan P, Aprison MH (1955) Cholinesterase activity in cytoplasmatic particles from rabbit brain. Fed Proc 14:106–107

    Google Scholar 

  • Silver A (1974) The biology of cholinesterases. North Holland, Amsterdam

    Google Scholar 

  • Skangiel-Kramska J, Niemierko S (1975) Soluble and particle bound acetylcholinesterase and its isoenzymes in peripheral nerves. J Neurochem 24:1135–1141

    Google Scholar 

  • Tennyson VM, Brzin M, Kremzner LT (1973) Acetylcholinesterase activity in the muscle satellite cell of the fetal rabbit. An electron microscopic-cytochemical and biochemical study. J. Histochem Cytochem 21:634–652

    Google Scholar 

  • Tsuji S (1974) On the chemical basis of thiocholine methods for demonstration of acetylcholinesterase activities. Histochemistry 42:99–110

    Google Scholar 

  • Tsuji S (1986) Histochemical demonstration of membrane-bound acetylcholinesterase: a methodological consideration on Koelle's copper thiocholine reaction and on Karnovsky's copper ferrocyanide reaction. Acta Histochem Suppl 33 (in press)

  • Tsuji S, Larabi Y (1983) A modification of thiocholine-ferricyanide method of Karnovsky and Roots for localization of acetylcholinesterase activity without interference by Koelle's copper thiocholine iodide precipitate. Histochemistry 78:317–323

    Google Scholar 

  • Wenk H, Krug H, Flechtner AM (1973) Eine mikrospektrophotometrische Methode zur quantitativen Aktivitätsbestimmung von Acetylcholinesterase an Gewebeschnitten. Acta Histochem 45:37–60

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

In honour of Prof. P. van Duijn

Rights and permissions

Reprints and permissions

About this article

Cite this article

Andrä, J., Lojda, Z. A histochemical method for the demonstration of acetylcholinesterase activity using semipermeable membranes. Histochemistry 84, 575–579 (1986). https://doi.org/10.1007/BF00482994

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00482994

Keywords

Navigation