牦牛Cyclin D1基因的克隆及IGF-I对其在睾丸支持细胞表达的影响

doi:10.11843/j.issn.0366-6964.2018.02.010

Abstract

Abstract:

The aim of this study was to clone the CDS of yak (Bos grunniens) Cyclin D1 gene, to analyse its biological characteristics, and to research the effects of insulin-like growth factor-I (IGF-I) on Cyclin D1 gene and protein expression in Sertoli cell (SC). The testicular samples were collected from 5-to 8-month-old healthy yaks for isolation and culture of SC. The Cyclin D1 gene was cloned by RT-PCR and its biological characteristics were analyzed by ORF Finder, MEGA7.0 and DNAMAN. The expression of Cyclin D1 protein in SC was localized by immunofluorescence staining. IGF-I was added into the culture medium of SC in the concentration of 0(control), 25, 50, 100, 150 ng·mL^-1, and then the relative expressions of Cyclin D1 mRNA and protein were detected by real-time quantitative PCR (qRT-PCR) and Western blotting, respectively. The results were as follows:1) Yak Cyclin D1 gene (GenBank accession number:KY 420723) had the high homology with other species. 2) Cyclin D1 protein belonged to nuclear protein because of high fluorescence intensity on cell nucleus. 3) The level of Cyclin D1 mRNA in 25 and 150 ng·mL^-1 groups were not significantly different with the control group (P>0.05), while 50 and 100 ng·mL^-1 groups were significantly higher than other groups (P<0.05); the difference of Cyclin D1 protein expression between 100 ng·mL^-1 group and the control group was not significantly different (P>0.05), however, a significant difference existed between 25, 50, 150 ng·mL^-1 groups and control group (P<0.05); the relative expression of Cyclin D1 mRNA and protein were all reached the highest levels when IGF-I was 50 ng·mL^-1, which were 1.65 and 1.20 times higher than that of the control group, respectively. The results suggested that Cyclin D1 gene was highly conserved during evolutionary process; IGF-I could regulate the expression of Cyclin D1 in SC and its optimal concentration was 50 ng·mL^-1.

CLC Number:

S823.8⁺5.2

WANG Ya-ying, PAN Yang-yang, YE Xiao-lin, LI Gu-yue, HE Hong-hong, WANG Meng, FAN Jiang-feng, CUI Yan, YU Si-jiu. Cloning of Yak Cyclin D1 Gene and the Effects of IGF-I on Its Expression in Sertoli Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 310-317.

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Cloning of Yak Cyclin D1 Gene and the Effects of IGF-I on Its Expression in Sertoli Cells

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