Developmental competence of mature yak vitrified–warmed oocytes is enhanced by IGF-I via modulation of CIRP during in vitro maturation |
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| Institution: | 1. Gansu Province Livestock Embryo Engineering Research Center, College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China;2. University of South Bohemia in ?eské Budějovice, Faculty of Fisheries and Protection of Waters, South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses, Research Institute of Fish Culture and Hydrobiology, Záti?í 728/II, 389 25 Vodňany, Czech Republic;1. Smithsonian Conservation Biology Institute (SCBI) and Hawaii Institute of Marine Biology (HIMB), Kaneohe, HI, USA;2. Zebrafish International Research Center, University of Oregon, Eugene, OR, USA;4. Aquatic Germplasm and Genetic Resources Center, Louisiana State University Agricultural Center (LSUAC), Baton Rouge, LA, USA;1. Agrotecnio Center, University of Lleida, 25198, Lleida, Spain;2. Transfer in Bovine Reproduction SLu, 22300, Barbastro, Spain;3. Sidney Sussex College, University of Cambridge, England, UK;4. Ladfield, Oxnam, Jedburgh, TD8 6RJ, Scotland, UK;1. College of Animal Science and Technology, Shihezi University, Shihezi, 832003, China;2. Key Laboratory of Genetics Breeding and Reproduction of Grass Feeding Livestock, Ministry of Agriculture and Rural affairs, Urumqi, 830000, PR China;3. Inner Mongolia Sino Sheep Technology Co. Ulanchap, 011800, China;4. HouBo College of Xinjiang Medical University, Karamay, 834000, China |
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| Abstract: | The objective of this study was to investigate whether developmental competence of mature vitrified–warmed yak (Bos grunniens) oocytes can be enhanced by supplemented insulin-like growth factor I (IGF-1) during in vitro maturation (IVM), and its relationship with the expression of cold-inducible RNA-binding protein (CIRP). In experiment 1, immature yak oocytes were divided into four groups, and IVM supplemented with 0, 50, 100 and 200 ng/mL IGF-1 was evaluated; the mRNA and protein expression levels of CIRP in mature oocytes in the four groups were evaluated using quantitative real-time PCR and western blotting analyses. In experiment 2, the mature yak oocytes in the four groups were cryopreserved using the Cryotop (CT) method, followed by chemical activation and in vitro culture for two days and eight days to determine cleavage, blastocyst rates, and total cell number in the blastocysts. Mature yak oocytes without vitrification served as a control group. The outcomes were as following: (1) the expression of CIRP in the matured oocytes was up-regulated in the IGF-1 groups and was highest expression was observed in the 100 ng/mL IGF-1 treatment group. (2) In the vitrified–warmed groups, the rates of cleavage and blastocyst were also highest in the 100 ng/mL IGF-1 treatment group (81.04 ± 1.06%% and 32.16 ± 1.01%), which were close to the rates observed in groups without vitrification (83.25 ± 0.85% and 32.54 ± 0.34%). The rates of cleavage and blastocyst in the other vitrified–warmed groups were 70.92 ± 1.32% and 27.33 ± 1.31% (0 ng/mL); 72.73 ± 0.74% and 29.41 ± 0.84% (50 ng/mL); 72.43 ± 0.61% and 27.61 ± 0.59% (200 ng/mL), respectively. There was no significant difference in the total cell number per blastocysts between the vitrified–warmed groups and group without vitrification. Thus, we conclude that the enhancement in developmental competence of mature yak vitrified–warmed oocytes after the addition of IGF-1 during IVM might result from the regulation of CIRP expression in mature yak oocytes prior to vitrification. |
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| Keywords: | Yak Mature oocytes Developmental competence CIRP IGF-1 Vitrification |
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