精子数量是影响男性生育的重要因素。我们前期发现倒班工人等人群睡眠行为的改变可能导致精子数量下降，其机制尚不清楚。近期有文献报道睡眠行为变化干扰昼夜节律关键基因Clock等的表达，而后者参与调控精子发生。但睡眠行为变化是否通过干扰昼夜节律关键基因表达而导致精子损害尚未证实。本课题拟开展以下研究：1）通过控制光照-黑暗周期和喂食时间的方法操纵小鼠睡眠行为改变，对昼夜节律关键基因Clock等表达造成干扰2）在此基础上通过生精上皮时相和精子数量等指标评价睡眠行为改变造成的精子损害并确认其发生阶段3）从反向和正向两方面调控昼夜节律基因表达并观察效应：在Clock基因敲除小鼠中观察睡眠行为变化是否仍导致精子损害；在野生型小鼠中以RNAi缓解Clock等表达所受干扰并观察精子损害是否改善，以明确昼夜节律关键基因在睡眠行为变化致精子损害中的作用，为阐明精子损害机制、维护男性生殖健康和行为干预提供理论依据。

Sperm number is key index of male fertility. Our previous researches in shift workers and other population suggested that the frequent change of sleep behaviors was associated with decreased sperm number in the population, but the underlying mechanism remains unknown. There are evidences suggesting that sleep behavior influences the expression of circadian genes including Clock, which were involved in the spermatogenesis process. But it is not validated whether the change of sleep behaviors induced sperm damage by disrupting the circadian genes expression. The plan of the present study is: 1)To modify the sleep of animals by controlling the light/dark cycle and feeding schedule, so that expression of circadian genes including Clock in the testes will be disrupted. 2) To measure the sperm biomarkers in testes and epididymis and evaluate the damage of spermatozoa as well as the stage in which the damage happens. 3) To modify the circadian gene expression in different directions and investigate whether the sleep-related sperm damage was influenced: to investigate the sperm biomarkers in the Clock knockout animals whose peripheral circadian rhythm are abolished in order to see whether the sleep change still induce sperm damage; and to attenuate the disruption of the circadian genes including Clock in the wild type animals by RNAi technique and investigate whether the spermatozoa damage was relieved, with expectation of clarifying the role of circadian genes in the sleep-related spermatozoa damage. This study will help to understand the mechanism of spermatozoa damage and to keep the male fertility in the population.