The circadian Clock gene regulates acrosin activity of sperm through serine protease inhibitor A3K.
Our previous study found that CLOCK knockdown in the testes of male mice led to a reduced fertility, which might be associated with the lower acrosin activity. In this present study, we examined the differential expression in proteins of CLOCK knockdown sperm. Clock gene expression was knocked down in cells to confirm those differentially expressions and serine protease inhibitor SERPINA3K was identified as a potential target. The up-regulated SERPINA3K revealed an inverse relationship with Clock knockdown. Direct treatment of normal sperm with recombinant SERPINA3K protein inhibited the acrosin activity and reduced in vitro fertilization rate. The luciferase reporter gene assay showed that the down-regulated of Clock gene could activate the Serpina3k promoter, but this activation was not affected by the mutation of E-box core sequence. Co-IP demonstrated a natural interaction between SERPIAN3K and RORs (α and β). Taken together, these results demonstrated that SERPINA3K is involved in the Clock gene-mediated male fertility by regulating acrosin activity and provide the first evidence that SERPINA3K could be regulated by Clock gene via retinoic acid-related orphan receptor response elements.
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China.
Shuting
S
Cheng
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China Reproductive Medical Center of The Second Affiliated Hospital, Chengdu University of Traditional Chinese Medicine, Chengdu 610041, PR China.
Xin
X
Liang
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China.
Yuhui
Y
Wang
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China.
Zhou
Z
Jiang
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China.
Yanyou
Y
Liu
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China.
Wang
W
Hou
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China.
Shiping
S
Li
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China Department of Microbiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China.
Jing
J
Zhang
Health Ministry Key Laboratory of Chronobiology, College of Basic Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, PR China wangzhengrong@126.com.
Zhengrong
Z
Wang
D000176:Acrosin / Q000096:biosynthesis*; D000818:Animals; D056926:CLOCK Proteins / Q000172:deficiency / Q000378:metabolism*; D057906:Circadian Clocks; D005260:Female; D005298:Fertility; D005786:Gene Expression Regulation; D055786:Gene Knockout Techniques; D008297:Male; D008813:Mice, Inbred ICR; D015843:Serpins / Q000378:metabolism*; D013094:Spermatozoa / Q000502:physiology*
C490400:Serpina3k protein, mouse; D015843:Serpins; D056926:CLOCK Proteins; C538944:Clock protein, mouse; D000176:Acrosin
Clock gene; SERPINA3K; acrosin activity; male reproduction; serine protease inhibitor