A non‐invasive simple method for measurement of urinary excretion of melatonin in undisturbed mice

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Abstract
Melatonin is a hormone involved in neuroendocrine responses; its plasma concentrations display a circadian pattern which is modified by stress. Studies for determining the effects of stressors on melatonin levels in laboratory animals present the difficulty that the procedures for blood sampling are by themselves potential stressors capable of influencing the levels of the hormone measured. A simple non-stressful method for measuring urinary excretion of melatonin has been consequently developed. The method is applicable to single undisturbed mice kept in conventional cages, and consists of urine collection on chromatographic paper followed by extraction and melatonin assay by radioimmunoassay. The use of this method with BD2F1 mice indicates nocturnal excretion of melatonin significantly higher than during the day; nighttime melatonin levels were shown to be suppressed by constant illumination. A significant increase in nocturnal melatonin excretion was caused by the application of rotational stress applied as a mild experimental stressor.