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| Expression of COL1A1 Gene in the Gonad Axis of Guizhou Black Goat (Capra hircus) and Its Effect on Lambing Related Genes |
| ZHOU Ming-Shuai, WEN Xiao-Yan, ZHANG Yan, LI Yong, XU Chang, YANG Xiu-Yuan, ZHAO Jia-Fu* |
| Key Laboratory of Plateau Mountain Animal Genetics, Breeding and Reproduction, Ministry of Education/College of Animal Science, Guizhou University, Guiyang 550025, China |
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Abstract Collagen typeⅠα1 chain (COL1A1) gene as an important component of typeⅠcollagen, plays a role in regulating follicular development and ovarian cell proliferation, and is closely related to animal reproductive traits. In order to explore the relationship between COL1A1 gene and fertility of Guizhou black goats (Capra hircus) and the effects of different expression levels on related genes in lambing, in this study, gonad axis tissues (uterus, ovary, pituitary, hypothalamus, fallopian tube) of monotocous and polytocous Guizhou black goats were collected. The expression of COL1A1 gene and its protein in the gonadal axis of monotocous and polytocous Guizhou black goats were detected by qPCR, and then ovarian granulosa cells were isolated and cultured. The interference vector of the target gene was constructed by RNA interference technology and transfected into ovarian granulosa cells. The expression level of COL1A1 gene and its protein was detected by qPCR and Western blot to screen the best interference group. qPCR was used to detect the effects of COL1A1 gene interference on the expression of lambing related genes bone morphogenetic protein 15 (BMP15), bone morphogenetic protein 1B (BMPR-1B), growth differentiation factor 9 (GDF9) and follicle stimulating hormone β-subunit (FSHβ). The results showed that COL1A1 gene was expressed in various tissues of Guizhou black goats. The expression level in ovary and hypothalamus of polytocous group was the highest, and was extremely significantly higher than that of monotocous Guizhou black goats group (P<0.01), significantly higher in pituitary gland and oviduct than in monotocous group (P<0.05), there was no significant difference in utero; Immunofluorescence results showed that the cells isolated and cultured in the experiment were ovarian granulosa cells. qPCR and Western blot results showed that shRNA-COL1A1-2 was the best interference group. After inhibiting the expression of COL1A1 gene, qPCR results showed that the expressions of lambing related genes BMP15, BMPR-1B, GDF9 and FSHβ were significantly lower than those of shRNA-NC group (P<0.01). This study revealed the difference of COL1A1 gene expression in gonad axis of monotocous and polytocous Guizhou black goat. The high expression of COL1A1 gene in the ovarian tissue of polytocous lambs group and the suppression of lambing related genes by silencing COL1A1 gene indicated that COL1A1 gene was closely related to ovarian development and follicle growth. It can be used as a candidate gene for the breeding of Guizhou black goat.
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Received: 24 December 2021
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Corresponding Authors:
* jfzhao@gzu.edu.cn
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