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.
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References
Andrä J, Duijn P van (1985) Quantitative aspects of cytochemical methods for acetylcholinesterase studied with a cytochemical model system. Histochemistry 83:443–449
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
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
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
Giacobini E (1960) The intracellular localization of cholinesterases. J Histochem Cytochem 8:419–424
Hollunger EG, Niklasson BH (1973) The release and molecular state of mammalian brain acetylcholinesterase. J Neurochem 20:821–836
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
Hüther G, Luppa H (1979) The multiple forms of brain acetylcholinesterase III. Implications for the histochemical demonstration of acetylcholinesterase. Histochemistry 63:115–121
Karnovsky MJ, Roots L (1964) A “direct coloring” thiocholine method for cholinesterase. J Histochem Cytochem 12:219–221
Koelle GB, Friedenwald JS (1949) A histochemical method for localizing cholinesterase activity. Proc Soc Exp Biol (NY) 70:617–622
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
Lawler H (1964) The preparation of a soluble acetylcholinesterase from brain. Biochim Biophys Acta 81:280–288
Lewis PR (1961) The effect of varying the conditions in the Koelle technique. Bibl Anat 2:11–20
Lewis PR, Knight DP (1977) Staining methods for sectioned material. North Holland, Amsterdam
Lojda Z, Gossrau R, Schiebler TH (1979) Enzyme histochemistry. A laboratory manual. Springer, Berlin Heidelberg New York
Lukáš Z, Fialová Z (1976) The possibilities and limitation of membrane methods for the histochemical demonstration of cholinesterases. Histochemistry 46:245–248
Massoulié J, Bon S (1982) The molecular forms of cholinesterase and acetylcholinesterase in vertebrates. Annu Rev Neurosci 5:57–106
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
Nathan P, Aprison MH (1955) Cholinesterase activity in cytoplasmatic particles from rabbit brain. Fed Proc 14:106–107
Silver A (1974) The biology of cholinesterases. North Holland, Amsterdam
Skangiel-Kramska J, Niemierko S (1975) Soluble and particle bound acetylcholinesterase and its isoenzymes in peripheral nerves. J Neurochem 24:1135–1141
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
Tsuji S (1974) On the chemical basis of thiocholine methods for demonstration of acetylcholinesterase activities. Histochemistry 42:99–110
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
Wenk H, Krug H, Flechtner AM (1973) Eine mikrospektrophotometrische Methode zur quantitativen Aktivitätsbestimmung von Acetylcholinesterase an Gewebeschnitten. Acta Histochem 45:37–60
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In honour of Prof. P. van Duijn
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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
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DOI: https://doi.org/10.1007/BF00482994