Abstract：Cryptochrome 1 (CRY1) gene is a member of mammalian clock genes. Its expressed protein is both a photoreceptor and a part of the circadian clock. It can be delivered to the effector via the humoral and neural pathways to regulate mammalian physiology, biochemistry and behavior. In this study, the samples (the hypothalamus, pituitary, pineal and testicular tissues) of yak were collected, and the sequence of CRY1 coding region was cloned and analyzed. The hypothalamus, pituitary, pineal gland and testes of Qinghai Yak (Bos grunniens) were collected. The sequence of CRY1 coding region was cloned by RT-PCR. The expression of CRY1 in adult male yak reproductive axis and yak testes of different ages (30 days, 2 years, 4 years and 6 years) was analyzed by Real-time fluorescent quantitative PCR（qRT-PCR）, immunoblotting and immunohistochemical staining. The results showed that the CRY1 gene CDS was 1 712 bp in length and the ORF was 1 257 bp, encoding a total of 418 amino acid residues, no transmembrane domain, and the encoded protein was a soluble protein. Tissue expression and localization revealed that CRY1 mRNA and protein were expressed in the hypothalamus, pituitary, testis and pineal tissues of yak, and the expression level in pineal tissue was significantly higher than other tissues (P<0.01). Immunohistochemical staining showed positive expression in the hypothalamic paraventricular nucleus, pituitary cells, testicular spermatogenic cells, pineal cells and glial cells. The expression of CRY1 mRNA and protein in the testis of 30 days old was the lowest in the testis tissues of different age groups, and the expression level gradually increased with age.different age groups was the lowest in the 30-day testis. The results showed that the CRY1 gene was relatively more conservative in animal evolution and was expressed in the male yak gland and the yak testes of different ages, suggesting that it might play an important role in the reproductive process of yak. The above results provided the basis for the biological function of the yak biorhythm gene CRY1 and its related research on the regulation of yak reproductive physiology.
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