Abstract
Maternal protein components change markedly during mammalian oogenesis. Many of these proteins have yet to be characterized and verified. In this study, a proteomics approach was used to evaluate changes in proteins during oogenesis in the Swamp Buffalo (Bubalus bubalis). Proteins from 500 immature oocytes and 500 in vitro matured oocytes were subjected to two-dimensional electrophoresis, and more than 400 spots were detected. Image analysis indicated that 17 proteins were differentially expressed between the two groups. Eight proteins were identified by mass spectrometry. In mature oocytes, three proteins were down-regulated: major vault protein (MVP), N-acetyllactosaminide β-1,6-N-acetylglucosaminyl-transferase (GCNT-2), and gem-associated protein (GEMIN)8, whereas five other proteins, heat shock protein (HSP)60, Ras-responsive element-binding protein 1 (RREB-1), heat shock cognate 71 kDa protein (HSC71), hemoglobin subunit α (HBA), and BMP-2-inducible protein kinase (BMP-2K), were up-regulated. The expression profiles of HSP60 and GEMIN8 were further verified by Western blotting. The changes in HSP60 protein expression demonstrate the increasing need for mitochondrial protein importation to facilitate macromolecular assembly during oocyte maturation. The down-regulation of GEMIN8 production implies that RNA splicing is impaired in mature oocytes.
Highlights
Mammalian oocytes undergo a series of regulated events during follicular development in the ovary, from primordial follicles to ovulation
Some well-known oocyte proteins, such as Stella [3], Zar1 [4], and Mater [5], play important roles in zygote activation. Many aspects of these mechanisms have been studied in order to understand the molecular foundations of oocyte maturation, and proteomic technology has been used for some species
We identified and verified some target proteins that might be involved in oocyte maturation in swamp buffalo
Summary
Mammalian oocytes undergo a series of regulated events during follicular development in the ovary, from primordial follicles to ovulation. Some well-known oocyte proteins, such as Stella [3], Zar1 [4], and Mater [5], play important roles in zygote activation Many aspects of these mechanisms have been studied in order to understand the molecular foundations of oocyte maturation, and proteomic technology has been used for some species. Zhang et al (2009) [10] identified a total of 625 different proteins from 2700 mature oocytes, lacking zona pellucidae, using SDS–PAGE combined with high performance liquid chromatography (HPLC). They screened 76 maternal proteins with high levels of mRNA expression, both in oocytes and zygotes. More than 5000 distinct proteins were quantified including some important stem cell reprogramming markers
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