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A diurnal oscillation of acetylcholine concentrations in rat brain has been demonstrated by gas chromatography. Peak concentrations occur at 2 hours of light, and a trough is seen at 6 hours of darkness. This pattern is observed only in grouped rats, and emerges after at least 18 days of prior conditioning in an environment with controlled light, humidity, and temperature.
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Hanin et al. (1970) studied this question.
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