玻璃化冷冻对猪孤雌激活囊胚中细胞凋亡的影响

doi:10.11843/j.issn.0366-6964.2017.02.007

Abstract

Abstract:

The objective of this study was to evaluate the effect of vitrification on cell apoptotic levels of porcine parthenogenetic blastocysts. The blastocoel recovery, mitochondrial membrane potential (ΔΨm), early apoptotic levels and the activities of several caspases of frozen thawed blastocysts were measured, and the mRNA expression levels of apoptosis related genes involved in different apoptotic pathways were also measured. The results showed that the rate of re-expanded blastocysts (31.03%) and total cells (31.81) of vitrified blastocysts were significantly lower than those of fresh group (100.00% and 38.17, P<0.05). The mean mitochondrial ΔΨm of vitrification group was 0.46, which was significantly lower than that of fresh group (1.02, P＜0.05). TUNEL assay revealed a significantly higher rate of DNA fragmentation in the cells of vitrified blastocysts (10.13%), which was significantly higher than that of fresh group (5.92%，P＜0.05). This study also demonstrated that vitrification caused a significant increasing of Pan-caspase, Caspase-3, Caspase-8 and Caspase-9 activities in vitrified group (20.65, 20.60, 17.58 and 19.88) comparing with fresh group (7.41, 6.46, 5.47 and 6.32, P＜0.05). qRT-PCR results showed that the relative expression levels of pro-apoptotic genes (Caspase-8, Caspase-9, TNF-α) were up-regulated and the relative expression of anti-apoptotic genes (Bcl-2 and SOD-1) were down-regulated greatly in vitrified group (P＜0.05). This study concluded that the in vitro developmental competence of blastocysts after vitrification was reduced by the decreasing of mitochondrial function, and the increasing of apoptotic levels were mediated by both death receptor and mitochondria apoptotic pathways.

CLC Number:

S828.2

CHEN Ya-ning, WU Cai-feng, DAI Jian-jun, ZHANG Shu-shan, NIU Ying-fang, ZHANG De-fu. Effect of Vitrification on Cell Apoptotic Levels of Porcine Parthenogenetic Blastocysts[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 243-251.

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Effect of Vitrification on Cell Apoptotic Levels of Porcine Parthenogenetic Blastocysts

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