基于高通量测序的犏牛囊胚玻璃化冷冻损伤机制研究

doi:10.11843/j.issn.0366-6964.2017.10.010

Abstract

Abstract:

This study was conducted to explore the molecular mechanism for vitrified-thawed damage of the cross-bred blastocysts of the yak by RNA-seq. The cross-bred blastocysts were derived from yak oocytes in vitro fertilized with cattle spermatozoa. Total RNA were extracted from these fresh blastocysts (FRB) and vitrified-thawed blastocysts (VTB) and cDNA was amplified via the Smart-seq2 method, and finally RNA libraries were constructed and sequenced. We obtained 51 099 116 and 54 192 358 clean reads from FRB and VTB, respectively, of which more than 80% were mapped to referenced genome. A total of 11 196 differentially expressed genes (DEGs) were detected, in which 7 570 were upregulated, and 3 626 were downregulated in VTB vs. FRB. The FRB and VTB had 49 016 and 64 352 alternative splicing, 16 681 and 224 750 putative SNPs, respectively. GO enrichment analysis of DEGs showed that they were enriched in molecular function, cellular component, and biological process. The KEGG analysis of DEGs showed that there were 318 pathways in VTB and FRB, of which 14 pathways were significantly enriched. We speculated that the coordination between Spliceosome, Ubiquitin mediated proteolysis and protein processing in endoplasmic reticulum pathway's etc. and the differential expression of Prp19, Prp4, CaM, PKA, Hsp70 and CCL2 might play an important role in the mechanism for vitrified-thawed damage. This is the first report to explore the mechanism for damage of VTB using high-throughput sequencing, which might serve as a key resource for improving embryo vitrification technology. The study also provided valuable information to study gene structure of cross-bred yak, and discover new genes related to vitrification damage of embryos.

CLC Number:

S823.8⁺5.2

ZHENG Jie, PU Si-ying, YANG Yuan-xiao, WANG Qin, YANG Rao-fen, ZI Xiang-dong. Exploring Mechanism for Vitrification Damage of the Cross-bred Blastocysts of the Yak via High-throughput Sequencing[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(10): 1871-1881.

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Exploring Mechanism for Vitrification Damage of the Cross-bred Blastocysts of the Yak via High-throughput Sequencing

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