Abstract
To investigate the protein profiling of buffalo oocytes at the germinal vesicle (GV) stage and metaphase II (MII) stage, an iTRAQ-based strategy was applied. A total of 3,763 proteins were identified, which representing the largest buffalo oocytes proteome dataset to date. Among these proteins identified, 173 proteins were differentially expressed in GV oocytes and competent MII oocytes, and 146 proteins were differentially abundant in competent and incompetent matured oocytes. Functional and KEGG pathway analysis revealed that the up-regulated proteins in competent MII oocytes were related to chromosome segregation, microtubule-based process, protein transport, oxidation reduction, ribosome, and oxidative phosphorylation, etc., in comparison with GV and incompetent MII oocytes. This is the first proteomic report on buffalo oocytes from different maturation stages and developmental competent status. These data will provide valuable information for understanding the molecular mechanism underlying buffalo oocyte maturation, and these proteins may potentially act as markers to predict developmental competence of buffalo oocyte during in vitro maturation.
Highlights
In vitro maturation (IVM) of mammalian oocytes is an important technique for animal embryo technology, which directly affects the efficiencies of in vitro fertilization (IVF), nuclear transfer and transgenic etc
To identify the differentially expressed proteins in buffalo oocytes before and after maturation, germinal vesicle (GV) and metaphase II (MII) stage oocytes were selected for quantitative proteomics analysis by Isobaric tags for relative and absolute quantitation (iTRAQ)
After separated by SDS-PAGE (Fig. 1C) and in-gel digested with trypsin, the peptides were labeled with iTRAQ regents
Summary
In vitro maturation (IVM) of mammalian oocytes is an important technique for animal embryo technology, which directly affects the efficiencies of in vitro fertilization (IVF), nuclear transfer and transgenic etc. In order to investigate the gene expression profile of oocytes during maturation process and identify differential expression genes/transcripts in oocyte at different developmental stages and with different competence, various approaches, such as genomics and transcriptomics have been performed in the last decade. The related studies were mainly focused on identifying protein expression profile of embryos at different developmental stages, and maternal proteins in oocytes. Quantification is performed via reporter ion intensity from the low mass range at MS/MS level, which improves the accuracy of quantitation. This method could simultaneously analyze up to eight different samples in one experiment
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