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Effect of vitrification on the mRNA transcriptome of bovine oocytes
Author(s) -
Wang N,
Li CY,
Zhu HB,
Hao HS,
Wang HY,
Yan CL,
Zhao SJ,
Du WH,
Wang D,
Liu Y,
Pang YW,
Zhao XM
Publication year - 2017
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.12942
Subject(s) - transcriptome , biology , gene , messenger rna , gene expression , microbiology and biotechnology , kegg , oocyte , embryo , genetics
Contents Vitrification has been shown to decrease the developmental capacity of mammalian oocytes, and this is closely associated with the abnormal mRNA expressions of vitrified oocytes. However, the effect of vitrification on transcriptional machinery of oocytes examined by RNA sequencing ( RNA ‐seq) has yet to be defined. In the present study, the mRNA transcriptomes of fresh and vitrified bovine oocytes were analysed by Smart‐seq2 with the differently expressed genes determined by DE seq2 (an adjusted p ‐value of .05 and a minimum fold change of 2). The differentially expressed mRNA s were then searched against the Gene Ontology ( GO ) and Genomes ( KEGG ) database. Finally, the mRNA expressions of 10 candidate genes were validated using quantitative real‐time PCR ( qRT ‐ PCR ). Approximately 12,000 genes were detected in each sample of fresh or vitrified oocytes. Of these, the expression levels of 102 genes differed significantly in vitrified groups: 12 genes mainly involved in cell cycle, fertilization and glucose metabolism were upregulated, and 90 genes mainly involved in mitochondria, ribosomal protein, cytoskeleton, transmembrane protein, cell cycle and calcium ions were downregulated. GO analysis showed that these genes were mainly enriched in terms of membrane‐bounded organelles, macromolecular complex, and intracellular part. The mRNA expression levels of 10 candidate genes selected randomly were in agreement with the results of the RNA ‐seq. In conclusion, our results showed that vitrification affected the mRNA transcriptome of bovine oocytes by downregulating genes, which contributed to the decreased developmental capacity of vitrified oocytes. Our findings will be useful in determining approaches to improve the efficiency of vitrified oocytes.