Abstract
The existence of an alcohol-soluble, inactive compex of acetylcholine in cerebral tissue could not be confirmed. Acidulated alcohol was found to be a satisfactory extracting fluid for use in the determination of total Ach in brain tissue. Values observed by this method in dog, rabbit and rat brain are reported. Hypoxia did not change the total Ach content of dog brain frozen in situ. The Ach in excised cerebral tissue decreases slowly with time. After freezing and thawing, the decrease is very rapid. No decrease occurs if the tissue is frozen, ground, and subjected to alcoholic extraction while in the frozen state. Both synthesis and destruction of Ach were encountered in attempts to measure “free” and “bound” Ach in aqueous extracts of brain. The synthesis could be inhibited by addition of cupric chloride, but the destruction was not completely prevented by eserine. The tissue-bound Ach is largely released by thorough mechanical disintegration as well as by freezing and thawing. The ease with which this “complex” is disrupted by physical means makes the existence of a chemical combination appear improbable. It is concluded that measurements of “free” and “bound” Ach by aqueous extraction do not represent the state of affairs in the living brain.
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