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| Expression and Localization Analysis of AURKA in HPGA of Male Mouse (Mus musculus) and Effect of Its Knockdown on DHT Anabolism |
| ZHAO Cai-Ying1, LI Yi-Na2, WANG Qi2, GAN Ze2, ZHANG Yong1, 2, MA You-Ji3, ZHANG Quan-Wei1, *, ZHAO Xing-Xu1, 2, * |
1 College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
2 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
3 College of Animal Science and Technology, Gansu Agriculture University, Lanzhou 730070, China |
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Abstract Aurora kinase A (AURKA), one of the protein kinases, regulates mitotic protein kinase in cell cycle. Previous studies showed AURKA might be involved in regulating the development and maturation of germ cells. AURKA is a target gene of androgen receptor (AR) and dihydrotestosterone (DHT) to promote sperm maturation by combining with intracellular AR, which is of great significance for the study of animal reproduction and diseases, but the function of AURKA remains unclear. In the present study, the expression level and localization of AURKA in hypothalamus, pituitary and testis in male mice with 8 weeks were analyzed using semi-quantitative PCR, Western blot (WB), hematoxylin-eosin staining (HE) and immunohistochemistry (IHC). The designed specific knock down sequence of AURKA was transfected into the primary cultured sertoli cells. The expressed variation of AURKA and 5α-reductase type 1 (SRD5A1) mRNA and proteins was detected using the real-time fluorescent quantitative PCR (qRT-PCR) and WB after transfected for 24 h. The concentration of DHT in sertoli cells treated with or without AURKA-siRNA was determined using the enzyme-linked immunosorbent assay (ELISA). The results showed that AURKA mRNA and protein were expressed in the hypothalamic-pituitary-gonadal axis (HPGA) tissues of male mice. Compared to the expression of AURKA in hypothalamus and pituitary tissues, the highest expression level of AURKA mRNA and protein presented in testis (P<0.01). The strongest immuno-positive protein of AURKA showed in sertoli cells and sperm. The expression level of AURKA mRNA and protein was down-regulated (P<0.05), whereas the expression level of SRD5A1 mRNA and protein was significantly up-regulated (P<0.05) and the the concentration of DHT increased (P<0.05) in adult mice sertoli cells 24 h after the interference with AURKA-siRNA. The results suggested the expression of SRD5A1 and DHT synthesis were negatively regulated by AURKA. This data provide some theoretical basis for understanding the function of AURKA in the reproductive processes of male animals.
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Received: 14 January 2020
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Corresponding Authors:
* , zhangqw@gsau.edu.cn; zhaoxx@gsau.edu.cn
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