Proteomics

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Human oocyte proteome and secretome

ABSTRACT: The proteome and secretome of human oocytes was studied by shotgun LCMSMS, starting from pools of 100 cells. Exploiting SP3, a novel technology for proteomic sample preparation using magnetic beads, we scaled down proteome analysis to single cells. Despite the low protein content of only ~100ng per cell, we consistently identified ~450 proteins from individual oocytes. When comparing proteomes of individual oocytes at the germinal vesicle (GV) and metaphase II (MII) stage by label-free quantification, we found several proteins preferentially expressed in matuare and immature cells. This study demonstrates that an innovative proteomics workflow facilitates analysis of single human oocytes to investigate human oocyte biology and pre-implantation development. The approach presented here paves the way for quantitative proteomics in other quantity-limited tissues and cell types.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Oocyte

SUBMITTER: Jeroen Krijgsveld  

LAB HEAD: Jeroen Krijgsveld

PROVIDER: PXD003691 | Pride | 2016-05-26

REPOSITORIES: Pride

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Dataset's files

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Action DRS
100oocytesmature-immature_rawfiles.zip Other
100oocytesmature-immature_txtfiles.zip Other
10oocytesdimethyllabeling_rawfiles.zip Other
10oocytesdimethyllabeling_txtfiles.zip Other
secretome_rawfiles.zip Other
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Publications

Identification of Maturation-Specific Proteins by Single-Cell Proteomics of Human Oocytes.

Virant-Klun Irma I   Leicht Stefan S   Hughes Christopher C   Krijgsveld Jeroen J  

Molecular & cellular proteomics : MCP 20160523 8

Oocytes undergo a range of complex processes via oogenesis, maturation, fertilization, and early embryonic development, eventually giving rise to a fully functioning organism. To understand proteome composition and diversity during maturation of human oocytes, here we have addressed crucial aspects of oocyte collection and proteome analysis, resulting in the first proteome and secretome maps of human oocytes. Starting from 100 oocytes collected via a novel serum-free hanging drop culture system,  ...[more]

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