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      2. PGF2α信號對水牛卵母細胞和早期胚胎發育的影響
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        國家科技重大專項(2018ZX08008-01B); “十三五”國家重點研發計劃項目(2018YFD0501605); 中央高?;A研究經費項目(2662016QD038)


        Effect of PGF2α signal on oocyte and early embryo development in buffalo
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          摘要:

          為探究前列腺素F2α(PGF2α)信號對水牛繁殖能力的影響,利用免疫熒光染色和實時熒光定量PCR檢測PGF2α受體(PTGFR)在水牛卵母細胞和胚胎早期發育過程中的表達規律;并在卵母細胞體外成熟液、胚胎培養發育液中分別添加不同濃度的PTGFR抑制劑AL-8810,檢測其對水牛卵母細胞成熟效率以及胚胎發育效率的影響。結果顯示,PTGFR蛋白在水牛卵母細胞及胚胎中廣泛分布,在4-16細胞期胚胎階段,PTGFR基因表達水平顯著高于未受精的卵母細胞和桑椹胚;AL-8810對水牛卵母細胞核成熟效率無顯著影響,但高濃度處理組(1 600 nmol/L)顯著降低卵母細胞早期凋亡率,100和800 nmol/L的AL-8810可顯著提高體外16細胞期胚胎的發育效率,800 nmol/L組桑椹胚形成率最高。以上研究結果表明,抑制PGF2α信號可有效提高水牛卵母細胞體外成熟質量與早期胚胎發育潛力。

          Abstract:

          Buffalo has outstanding characteristics of stress resistance and adoptable to poor quality roughages and is mainly used for farming service in the world.Many of the economic traits of buffaloes have not been fully utilized and are therefore considered to be the most potential livestock for development.However,buffalo breeding methods of the world is still relatively traditional,and there are few modern scale farms,which greatly limits the development of buffalo industry and utilization of economic traits of buffalo.Compared with cattle,buffalo reproductive problems are mainly manifested in the low number of follicular reserves,insignificant estrous symptoms,low breeding efficiency,and easy loss of early embryos,which also directly limits the feeding scale of buffaloes.Therefore,with the help of modern reproductive technology (such as artificial insemination,superovulation-embryo transfer,in vitro embryo production,etc.),it is an important scientific and technological guarantee for the development of buffalo breeding industry to solve the prominent problems of buffalo reproduction in a short period of time.In vitro embryo production is an important reproductive technology,which can greatly expand the source of livestock embryos,and can improve the reproductive efficiency of livestock in a short period of time in combination with embryo transfers technology.The technical efficiency of in vitro embryo production mainly depends on in vitro oocyte maturation efficiency and early embryo development efficiency.How to optimize the methods of in vitro culture is the main focus of buffalo reproduction research.Reproductive hormones play a significant role in promoting oocyte maturation,sperm-egg interaction and embryo development and widely use in oocytes maturation and embryo culture systems,such as LH,FSH,E2 and P4.In addition,like an appropriate concentration of melatonin,vitamin C,resveratrol,sodium selenite can significantly improve the maturation quality of oocytes and promote development of embryos in vitro.Though a lot of work has been done to improve the in vitro culturing system of oocytes and embryos,yet the efficiency of both in vitro maturation of oocytes and embryonic development are significantly lower than in vivo.PGF2α signaling has a pivotal role in regulating animal reproduction and PGF2α receptor (PTGFR) is highly expressed in liver,ovary,uterus,adrenal glands,fat cells,thymocytes,pancreatic islets,platelets and red blood cells.Previously research has been reported that PGF2α plays an important role to inhibit the development of rabbit and mouse embryos in vitro.In dairy cows,the PGF2α receptor PTGFR is widely distributed in the reproductive organs,and the embryo quality and pregnancy rate are negatively correlated with the increase of the concentration of PGF2α in the uterine cavity.High level of PGF2α in the peripheral circulation of dairy cows has an inhibitory effect on embryonic development,resulting in a significant decrease of pregnancy rate of dairy cows.It’s speculated that inhibiting PGF2α signaling in vitro can improve oocyte maturation efficiency and early embryo development efficiency.Therefore,this study was aimed to investigate the effects of inhibiting PGF2α signaling on the maturation oocytes and development of early embryos in vitro.The expression patterns of PGF2α receptor (PTGFR) in buffalo oocytes and early embryo development were determined by immunofluorescence staining and real-time fluorescent quantitative PCR.Subsequently,the effects of PGF2α signaling on the maturation efficiency of oocytes and the efficiency of embryonic development in buffalo was studied by different concentrations of PTGFR inhibitor AL-8810 administration in the oocyte maturation medium and embryonic development medium.The results showed that PTGFR protein was widely distributed in buffalo oocytes and embryos,and the expression level of PTGFR gene was significantly higher in 4-16 cell stage than in unfertilized oocytes and morulae (P<0.05).Inhibiting PGF2α signaling by AL-8810 administration had no significant differences effect on the nuclear maturation efficiency of buffalo oocytes in different treatments(P>0.05),respectively.However,the early apoptosis rate of buffalo oocytes reduced significantly in high concentration treatment (1 600 nmol/L) (P<0.05).Embryonic development efficiency in 16 cell stage improved significantly in the treatments of concentration of 100 and 800 nmol/L,and the morula formation rate was the highest in the 800 nmol/L treatment.The above results indicated that inhibiting PGF2α signaling effectively improves in vitro maturation quality of oocytes and early embryo development potency in buffalo.The findings of this study about the above regulatory role of PGF2α signaling could provide new ideas for improving in vitro embryo production efficiency of buffalo.

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        冉早紅,何長久,張姝,劉清華,石六杰,王振,李翔. PGF2α信號對水牛卵母細胞和早期胚胎發育的影響[J].華中農業大學學報,2020,39(5):123-129

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        • 收稿日期:2020-03-31
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        • 在線發布日期: 2020-10-05
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