The pineal gland is a small neuroendocrine organ of the epithalamus, attached to the roof of the third ventricle. It secretes melatonin, linking a neural timing system to a chemical signal distributed beyond the brain. Its anatomical position is distinct from that of the nearby habenula.[ref]
A classic experiment located a strong daily rhythm at an enzymatic step in pineal metabolism. In 1970, D. C. Klein and J. L. Weller reported that rat pineal serotonin N-acetyltransferase activity was more than 15 times higher at night than during the day. The rhythm persisted in complete darkness and in blinded animals, whereas constant illumination suppressed it. Their experiments in isolated tissue also implicated norepinephrine in regulation of the enzyme activity.[ref]
The darkness control is particularly illuminating. A mechanism that merely switched on whenever light was absent would predict a continuously high output in constant darkness. Continued rhythmic variation instead supports an internally generated timing signal. It does not show that the pineal gland alone generates the organism's entire circadian rhythm: the measurement concerns one gland and one metabolic activity in an intact timing system. Likewise, the reported activity ratio is an enzyme-assay result, not a universal ratio of human blood melatonin concentrations.

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