蒙古羊双羔性状基因ADAMTS-1作用机制研究

doi:10.16431/j.cnki.1671-7236.2023.08.020

Abstract

Abstract: 【Objective】 Using single and twin Mongolian sheep as animal models,the mechanism of the main effect gene of double lambs containing type Ⅰ platelet-binding protein motif of disintegrin-like metalloproteinase (ADAMTS-1) was preliminarily explored to provide a theoretical basis for efficient breeding and related research in Mongolian sheep.【Method】 Twelve female Mongolian sheep with records of single and double births were selected,and use natural estrus methods to determine the estrus and interestrus periods of female Mongolian sheep through vaginal mucosal smears and male sheep crawling.The ovaries of Mongolian sheep during the stages of double lambing estrus (DE),double lambing dioestrum (DD),single lambing estrus (SE),and single lambing dioestrum (SD) were collected,and the morphological differences of ovarian antral follicles were observed using HE staining.Granulosa cells from the ovary of Mongolian sheep were cultured and the location of ADAMTS-1 expression was determined by immunofluorescence.The relative mRNA expression of ADAMTS-1,progesterone receptor (PGR),mitogen-activated protein kinase 4 (MAPK4),androgen receptor (AR),serine protease inhibitor 2 (SERPINE2) and multifunctional proteoglycan proteoglycan (Versican) were determined by Real-time quantitative PCR in different groups.And the relative protein expression of ADAMTS-1 precursor protein (pro-ADAMTS-1) and mature protein (MP-ADAMTS-1) in different groups were detected by Western blotting.【Result】 There were no apparent differences in the morphology of the luminal follicles within the ovarian tissue from single and twin Mongolian sheep,as determined by the results of HE staining,and ADAMTS-1 was detected on granulosa cells in the ovary of Mongolian sheep using immunofluorescence.The results of fluorescence PCR and Western blotting showed that the gene and protein of ADAMTS-1 in ovarian tissue of Mongolian sheep in DE group were significantly higher than those in the other three groups (P＜0.05),while there was no significant difference between SE and SD groups (P>0.05). The expression of AR and PGR genes in ovarian tissue of Mongolian sheep in SE group were the highest and significantly higher than those in the other 3 groups (P＜0.05),and that in DE group was also significantly higher than that in DD group (P＜0.05).Moreover,the expression of AR and PGR genes in ovary of single lambs at the same diestrus stage was significantly higher than in ovary of double lambs.The expression of MAPK4 gene in ovarian tissue of Mongolian sheep in SE group were the highest and significantly higher than that the other three groups (P＜0.05),while there was no significant difference among the other three groups (P＞0.05).The expression of Versican and SERPINE2 genes in ovarian tissue of Mongolian sheep in DD group were the highest and significantly higher than those in DE and SD groups (P＜0.05),these two factors in SE group were significantly higher than that in SD group (P＜0.05).【Conclusion】 ADAMTS-1 was stably expressed in granulosa cells of ovary in Mongolian sheep,and ADAMTS-1 had an important positive regulatory role on ovulation mechanism of double lamb trait in Mongolian sheep,while various other hormone receptors and intracellular key signaling genes were also closely related to follicle development and expulsion in ovary.

Key words: Mongolian sheep; ADAMTS-1; double lamb trait; ovary

CLC Number:

Q953.3

ZHAO Mengyao, CHEN Xiaoliang, LI Xiunan, SU Lede, TIAN Ying, JIAN Ruizhen, YANG Yanyan, LI Haijun. Study on the Mechanism of the Double Lamb Trait Gene ADAMTS-1 in Mongolian Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3229-3238.

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Study on the Mechanism of the Double Lamb Trait Gene ADAMTS-1 in Mongolian Sheep

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