Proteomic-based identification of maternal proteins in mature mouse oocytes

Abstract

BackgroundThe mature mouse oocyte contains the full complement of maternal proteins required for fertilization, reprogramming, zygotic gene activation (ZGA), and the early stages of embryogenesis. However, due to limitations of traditional proteomics strategies, only a few abundantly expressed proteins have yet been identified. Our laboratory applied a more effective strategy: one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D SDS-PAGE) and reverse-phase liquid chromatography tandem mass spectrometry (RP-LC-MS/MS) were employed to analyze the mature oocyte proteome in depth.ResultsUsing this high-performance proteomic approach, we successfully identified 625 different proteins from 2700 mature mouse oocytes lacking zona pellucidae. This is the largest catalog of mature mouse oocyte proteins compiled to date. According to their pattern of expression, we screened 76 maternal proteins with high levels of mRNA expression both in oocytes and fertilized eggs. Many well-known maternal effect proteins were included in this subset, including MATER and NPM2. In addition, our mouse oocyte proteome was compared with a recently published mouse embryonic stem cell (ESC) proteome and 371 overlapping proteins were identified.ConclusionThis proteomics analysis will be a valuable resource to aid in the characterization of important maternal proteins involved in oogenesis, fertilization, early embryonic development and in revealing their mechanisms of action.

Maternal age effect on mouse oocytes: new biological insight from proteomic analysis

“…Previous MS studies on mice analysed the proteome of mature oocytes , Zhang et al 2009) and processes of oocyte maturation (Vitale et al 2007, Cao et al 2012, Monti et al, 2013 in young donors, but not oocyte quality and composition during maternal ageing. As MS is not inherently quantitative, Wang et al (2010) relied on the simultaneous comparison of signal intensities between replica samples from young donors, in a so-called label-free approach that requires high numbers of oocytes.…”

Identification and Characterization of an Oocyte Factor Required for Porcine Nuclear Reprogramming

“…During the past decades, several approaches analyzing the protein composition of oocytes have been conducted to obtain the proteome signatures of Xenopus, bovine, pig, and mouse oocytes (5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15). Calvert et al (15) identified eight highly abundant heat shock proteins and related chaperones in the mature mouse egg by two-dimensional difference gel electrophoresis (DIGE).…”

Cytoplasmic Inheritance Redux

“…The number of mammalian genes that have been classified as maternal effect thus far is significantly less than the number of proteins one would expect to be necessary for pre-EGA embryonic development (Yurttas et al, 2010;Zhang et al, 2009). There are potentially many reasons for this, but I will consider five.…”