Proteome analysis of mouse oocyte pronuclei
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus Musculus (mouse)
SUBMITTER: Tomoya Kitajima
PROVIDER: PXD049709 | JPOST Repository | Sat Mar 01 00:00:00 GMT 2025
REPOSITORIES: jPOST
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Project description:Here we asked if oocyte proteomes are representative of the transcriptomes, how the abundance of specific genes’ mRNA and protein responds to maternal aging, and if oocyte aging presents the features characteristic of somatic aging. To address these questions on the proteomic level, we employed stable isotope labeling of amino acids in cell culture (SILAC; Geiger et al. 2011) as the method of choice, and performed a SILAC screen of mouse metaphase II (MII) oocytes superovulated at 3, 8+-1 and 58+-10 weeks of maternal age, which correspond to pre-puberty, mature age and climacterium, respectively. We used heavy F9 embryonic carcinoma (EC) cells as internal or "Spike-in" standard for the quantification of oocytes proteins because in contrast to the oocytes they can easily be cultured feeder-free, have stem cell properties and should harbor the majority of all oocyte proteins (although with different relative abundances). The SILAC screen was conducted in parallel with conventional microarray analysis to compare the concordance of protein and transcript levels in these oocytes. Associated microarray data have been deposited to NCBI GEO with accession number GSE42959.
2016-10-14 | PXD000512 | Pride
Project description:Oocyte-to-embryo transition plays a critical role in oocyte maturation and embryogenesis. It is a highly regulated process in part due to a transcription silenced period followed by zygotic genome activation. How transcriptome, translatome, and proteome interplay in this critical developmental window remains poorly understood. Utilizing a highly sensitive mass spectrometry, we obtained a high-quality proteome landscape spanning 10 stages, from the mouse full-grown oocyte (FGO) to blastocyst, using 100 oocytes/embryos at each stage. By integrative analysis with corresponding transcriptome and translatome, we found transcription and translation levels can not reflect protein abundance in most cases. From FGO to 4-cell embryos, proteomes are predominated by FGO-produced proteins, while the transcriptome and translatome are much more dynamic. FGO inherited proteins frequently persist after the corresponding transcripts are already downregulated or decayed. Improved concordance between protein and RNA is observed for genes starting translation only upon meiotic resumption (OET upregulated) or transcribed only in embryos, although the detected protein dynamics often lag behind transcription and translation. Concordance between protein and transcription/tranlation is associated with protein half-lives. Finally, a kinetic model well predicts protein dynamics when incorporating both the initial protein abundance in FGO and translation kinetics across developmental stages. In sum, our study reveals multilayer control of gene expression during oocyte maturation and embryogenesis.
2023-06-06 | GSE209648 | GEO
Project description:Mitochondria are essential organelles that provide energy for mammalian cells, with multiple functions such as signal transduction, cellular differentiation, and regulation of apoptosis. Compared with the mitochondria in somatic cells, oocyte mitochondria have an additional layer of importance since they are required for germ cell maturation, dysfunctions of which can lead to severe inherited disorders. Thus, a systematic proteomic profile of oocyte mitochondria is urgently needed for basic and clinical research, but has been hindered by the rarity of oocyte samples and technical challenges associated with capturing mitochondrial proteins from live oocytes. Here, in this work, using proximity labeling proteomics, we established a mitochondria-specific ascorbate peroxidase (APEX2) reaction in live GV staged oocytes of mouse and and explore oocyte mitochondrial proteomics.
2021-03-16 | PXD024777 |
Project description:PARP12 regulates mouse oocyte meiotic maturation
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Project description:Distinct transcriptome, translatome, and proteome reprogramming during the oocyte-to-embryo transition
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Project description:A long-standing question in developmental and reproductive biology is when the mammalian embryo becomes sufficiently distinct from its oocyte precursor. Myriads of studies examined the messenger RNAs that change during the oocyte-to-embryo transition, whereas proteins have been much less studied, in spite of their greater vicinity to phenotype. In the present study we modified the widely used embryo culture medium KSOM (PMID 12470333, PMID 10859270) to make it apt for our application. We replaced the serum albumin with polyvinylpyrrolidone and also replaced the natural Arginine and Lysine with their “heavy” isotopic variants Arginine 13C 15N and Lysine 13C 15N. Fertilized oocytes were retrieved from oviducts of gonadotropin-primed B6C3F1 females mated to CD1 males, and cultured at 37 degrees Celsius under 5% CO2 in KSOM containing 0.3 mM Arginine 13C 15N and 0.2 mM Lysine 13C 15N, which are the regular concentrations of these two amino acids in KSOM medium (PMID 12470333; PMID 10859270). After 4 days of culture, the embryos of the isotopic group had undergone blastocyst formation just like the control embryos cultured in normal medium. Samples of approx. 500 “heavy”-labeled blastocysts were collected zona-free and subjected to mass spectrometric analysis. The median labeling rate was 83%, ranging from 0% in proteins that did not incorporate any Arginine 13C 15N and Lysine 13C 15N, to 100% in proteins that were completely labeled. Our study demonstrates that a commonly used, chemically defined medium can be adapted for Stable Isotope Labeling by/with Amino acids in Cell culture (SILAC) and combined with high-resolution mass spectrometry, in a preimplantation embryo setting. This allows to tackle long-standing questions in developmental and reproductive biology, such as the identification of putative maternal (0% labeled), putative embryonic (100% labeled) or shared proteins in live mammalian embryos.
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Project description:To verify the Ribo-seq data of mouse oocyte, we performed MS/MS on mouse fully-grown oocytes. And the results show that our Ribo-seq data well reflect the proteomic dynamics in the fully grown oocytes.
2022-03-14 | PXD032229 | Pride
Project description:In this study, we applied 1D SDS-PAGE and RP-LC-MS/MS to investigate the proteins stored in GV mouse oocytes. This high-performance strategy allowed us to define a set of 1405 different mouse GV oocyte proteins. It is confirmed that this study will help us to understand the diverse biological processes occurring in mouse oocytes and during early embryo development. However, compared with proteomic analysis of other cells and tissues, such as embryonic stems from cells and liver, the proteins identified in mature mouse oocytes were limited. This was mainly due to the fact that oocytes obtained from each mouse were very limited. We believe that the catalog of maternal proteins presented in this article is a starting point and we anticipate that more researches on the oocyte proteome will deduce most of the maternal proteins.
2020-09-17 | PXD014314 | Pride
Project description:Within the ovarian follicle, mammalian oocytes acquire the necessary molecular machinery for successful fertilisation and embryonic development. In the absence of the follicle, oocytes matured in vitro (IVM) using traditional IVM show compromised developmental competence, resulting in poorer outcomes for patients. A novel in vitro system (CAPA-IVM) using c-type natriuretic peptide provides the oocyte with extended time to develop, with improved success rates. Here, we interrogate constituents of oocyte developmental competence by comparing the proteomic signature of mature mouse metaphase II (MII) cumulus-oocyte complexes (COCs) of differing intrinsic qualities, as matured by various maturation protocols. We employed label-free mass spectrometry to analyse the proteomes of mouse oocytes and cumulus cells separately. Proteomic profiling identified around 1600 proteins in oocytes and 3100 in cumulus cells across all three treatment groups (at least 2 peptides per protein). Differential expression analysis and pattern analysis collectively revealed a signature of proteins that were consistently differentially expressed between in vivo and in vitro oocyte maturation systems (log2FC of ± 1 and a p-value ≤ 0.05). These subsets of proteins were mapped to biological processes including eukaryotic translation, autophagy and endocytosis pathways within oocytes. Changes in reactive oxygen species detoxification and serine biosynthesis were observed in cumulus cells. Altogether, these findings provide new insights into the proteomic mechanisms underpinning oocyte developmental competence, with the potential to improve clinical oocyte diagnostics, expand on current clinical maturation systems and provide novel therapeutic options to infertile patients.
2026-08-24 | PXD073269 | Pride
Project description:The proteome changes were quantified in RNF114-/- mouse oocyte using TMT 6-plex by LC-MS/MS.
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