Sadeesh E M, Sikka P, Balhara A K, Balhara S
Division of Animal Physiology and Reproduction, ICAR-Central Institute for Research on Buffaloes, Hisar, 125001, India.
Division of Animal Genetics and Breeding, ICAR-Central Institute for Research on Buffaloes, Hisar, 125001, India.
Cytotechnology. 2016 Dec;68(6):2271-2285. doi: 10.1007/s10616-016-0022-y. Epub 2016 Sep 20.
The study examined the effects of different environmental stress on developmental competence and the relative abundance (RA) of various gene transcripts in oocytes and embryos of buffalo. Oocytes collected during cold period (CP) and hot period (HP) were matured, fertilized and cultured in vitro to blastocyst hatching stage. The mRNA expression patterns of genes implicated in developmental competence (OCT-4, IGF-2R and GDF-9), heat shock (HSP-70.1), oxidative stress (MnSOD), metabolism (GLUT-1), pro-apoptosis (BAX) and anti-apoptosis (BCL-2) were evaluated in immature and matured oocytes as well as in pre-implantation stage embryos. Oocytes reaching MII stage, cleavage rates, blastocyst yield and hatching rates increased (P < 0.05) during the CP. In MII oocytes and 2-cell embryos, the RA of OCT-4, IGF-2R, GDF-9, MnSOD and GLUT-1 decreased (P < 0.05) during the HP. In 4-cell embryos, the RA of OCT-4, IGF-2R and BCL-2 decreased (P < 0.05) in the HP, whereas GDF-9 increased (P < 0.05). In 8-to 16-cell embryos, the RA of OCT-4 and BCL-2 decreased (P < 0. 05) in the HP, whereas HSP-70.1 and BAX expression increased (P < 0.05). In morula and blastocyst, the RA of OCT-4, IGF-2R and MnSOD decreased (P < 0.05) during the HP, whereas HSP-70.1 was increased (P < 0.05). In conclusion, deleterious seasonal effects induced at the GV-stage carry-over to subsequent embryonic developmental stages and compromise oocyte developmental competence and quality of developed blastocysts.
本研究考察了不同环境应激对水牛卵母细胞和胚胎发育能力以及各种基因转录本相对丰度(RA)的影响。在寒冷期(CP)和炎热期(HP)采集的卵母细胞经体外成熟、受精并培养至囊胚孵化阶段。对参与发育能力(OCT-4、IGF-2R和GDF-9)、热休克(HSP-70.1)、氧化应激(MnSOD)、代谢(GLUT-1)、促凋亡(BAX)和抗凋亡(BCL-2)的基因在未成熟和成熟卵母细胞以及植入前阶段胚胎中的mRNA表达模式进行了评估。在CP期间,达到MII期的卵母细胞、卵裂率、囊胚产量和孵化率均有所增加(P<0.05)。在MII期卵母细胞和2细胞胚胎中,HP期间OCT-4、IGF-2R、GDF-9、MnSOD和GLUT-1的RA降低(P<0.05)。在4细胞胚胎中,HP期间OCT-4、IGF-2R和BCL-2的RA降低(P<0.05),而GDF-9增加(P<0.05)。在8至16细胞胚胎中,HP期间OCT-4和BCL-2的RA降低(P<0.05),而HSP-70.1和BAX表达增加(P<0.05)。在桑葚胚和囊胚中,HP期间OCT-4、IGF-2R和MnSOD的RA降低(P<0.05),而HSP-70.1增加(P<0.05)。总之,在GV期诱导的有害季节效应会延续到随后的胚胎发育阶段,并损害卵母细胞的发育能力和发育囊胚的质量。