Dataset Information

Differences in oocyte composition anticipate differences in embryo quality: a proteomic study in inbred mice

ABSTRACT: Current models of early development assign major roles to stochastic processes and epigenetic regulation, whose influence is considered to outweigh the genetic differences that exist between individuals. However, epigenetic agents are ultimately encoded by genes. It follows that the variable endowment of oocytes in these genes’ products could influence, prime or even predetermine developmental trajectories and features of embryos. As model system we used inbred strains of mice (129/Sv, C57Bl/6, C3H/HeN, DBA/2J). We examined if and in how far the composition of oocytes is conserved after we observed large variability in developmental competence and embryo quality across the four strains. Using state-of-the-art proteomics (SILAC LC-MS/MS) combined with transcriptomics (RNA deep sequencing) we quantified 2012 protein groups and 15205 transcripts present simultaneously in oocytes of all four strains. Proteome and transcriptome expression levels had little correlation with each other, whereby the endowment of oocytes in proteins with known or putative function in embryo development could not be predicted from mRNA abundance. Our data show that depending on the oocyte donor strain, resultant embryos have different qualities that correlate with the abundance of catalytic proteins, highlighting the importance of proteomic quantifications in modern embryology. Thus, different strains of mice should not be used interchangeably when tackling questions about oogenesis and early development, and a variety of strains should be studied to yield results of sufficient general validity.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Oocyte

SUBMITTER: Hannes Drexler

LAB HEAD: Hannes C. A. Drexler

PROVIDER: PXD001059 | Pride | 2015-01-28

REPOSITORIES: Pride

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			Action	DRS
	20130802_exp0550_Michele_B6_HPLC_01.raw	Raw
	20130802_exp0550_Michele_B6_HPLC_02.raw	Raw
	20130802_exp0550_Michele_B6_HPLC_03.raw	Raw
	20130802_exp0550_Michele_B6_HPLC_04.raw	Raw
	20130802_exp0550_Michele_B6_HPLC_05.raw	Raw

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Differences in embryo quality are associated with differences in oocyte composition: a proteomic study in inbred mice.

Pfeiffer Martin J MJ Taher Leila L Drexler Hannes H Suzuki Yutaka Y Makałowski Wojciech W Schwarzer Caroline C Wang Bingyuan B Fuellen Georg G Boiani Michele M

Proteomics 20150103 4

Current models of early mouse development assign roles to stochastic processes and epigenetic regulation, which are considered to be as influential as the genetic differences that exist between strains of the species Mus musculus. The aim of this study was to test whether mouse oocytes vary from each other in the abundance of gene products that could influence, prime, or even predetermine developmental trajectories and features of derivative embryos. Using the paradigm of inbred mouse strains, w ...[more]

PMID: 25367296

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Project description:The transformation of a fertilized oocyte into a multicellular embryo comprised of differentiated cells depends on the processing of genetic orders, first in the form of transcripts and then of proteins. Our current understanding of mouse development has greatly benefited from the analysis of transcripts as proxy for the proteins. However, the hexagonal-shaped free ribosomes that translate mRNAs into proteins are scarce during cleavage and do not become abundant until the morula-blastocyst stage. How accurate, then, is an understanding of mouse and more in general mammalian development that uses the analysis of transcripts as proxy for the proteins? This led us in this study to measure the proteome of seven preimplantation mouse stages (B6C3F1 x CD1), from oocyte to blastocyst, using 'spike-in' SILAC technology combined with high accuracy mass spectrometry (LTQ Orbitrap and Q-Exactive). The relative abundances of embryonic proteins were compared and contrasted with those of transcripts measured by RNA sequencing. A total of 6976 proteins were detected in at least the spike (precondition for determining the heavy/light peptide ratios in oocytes or embryos). Among these 6976 proteins, 4991 proteins were present in all developmental stages, and 1893 proteins were present in all replicates. Our tandem approach uncovered: 1) the different abundance profiles of proteome and transcriptome during the time of preimplantation development; 2) the different reciprocal associations of developmental stages (dendrograms) on the protein level as compared to the mRNA level; and 3) the different gene ontology terms overrepresented among the proteins that increase or decrease across adjacent stages, compared to mRNAs. In essence, the elaborate phasing of embryonic gene expression that has been known for mRNAs leaves an unsuspected footprint on the protein products of those mRNAs. This information facilitates the analysis of mammalian development in two important ways. First, it provides new insight into the regulation of the transition from the differentiated oocyte into the embryo, by identifying the subsets of proteins that accompany the passage from one embryonic stage to the next, which are likely composed of important regulators of the morphogenetic transitions. Second, our analysis provides a reference to determine the degree of ‘embryoness’ of entities produced via assisted reproductive technologies, cloning by somatic cell nuclear transfer, and even artificial gametes.