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Protein expression profiling in eggs and embryos from control and Eif4e1b knockout female mice.

ABSTRACT: We have identified eukaryotic translation initiation factor 4E family member 1B (Eif4e1b) as a regulator of maternal RNA translation, whose maternal deletion leads to 2-cell arrest phenotype in mouse embryos. eIF4E1b protein binds mRNA selectively in oocytes and controls their translation in early embryos. We aim to profile the global changes in protein expression in metaphase II (MII) eggs and zygotes collected from control or Eif4e1b knockout female mice. An encapsulated proteomic-sample processing protocol is used to perform this low-input mass spectrometry to quantify proteomic changes (Kulak et al. 2014/PMID 24497582).

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Embryo, Oocyte

SUBMITTER: David Anderson

LAB HEAD: Jurrien Dean

PROVIDER: PXD041583 | Pride | 2023-06-12

REPOSITORIES: Pride

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			Action	DRS
	allSpectra.HCD.ITMS.iso_0.apl	Other
	allSpectra.HCD.ITMS.iso_1.apl	Other
	allSpectra.HCD.ITMS.iso_10.apl	Other
	allSpectra.HCD.ITMS.iso_100.apl	Other
	allSpectra.HCD.ITMS.iso_101.apl	Other

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Publications

Germ cell-specific eIF4E1b regulates maternal mRNA translation to ensure zygotic genome activation.

Yang Guanghui G Xin Qiliang Q Feng Iris I Wu Di D Dean Jurrien J

Genes & development 20230531 9-10

Translation of maternal mRNAs is detected before transcription of zygotic genes and is essential for mammalian embryo development. How certain maternal mRNAs are selected for translation instead of degradation and how this burst of translation affects zygotic genome activation remain unknown. Using gene-edited mice, we document that the oocyte-specific eukaryotic translation initiation factor 4E family member 1b (eIF4E1b) is the regulator of maternal mRNA expression that ensures subsequent repro ...[more]

PMID: 37257918

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Identification of co-factors of eIF4E1b protein in mouse ovary.

Project description:We have identified eukaryotic translation initiation factor 4E family member 1B (Eif4e1b) as a regulator of maternal RNA translation, whose maternal deletion leads to 2-cell arrest phenotype in mouse embryos. eIF4E1b protein binds mRNA selectively in oocytes and controls their translation in early embryos. eIF4E1b expresses specifically in mouse ovary and we have generated a knock-in mouse line in which HA tag is conjugated to the C terminus of eIF4E1b. We aim to identify eIF4E1b co-factors using mass spectrometry after anit-HA immunoprecipitation (IP) using mouse ovaries.

2023-06-12 | PXD041622 | Pride

Association of maternal mRNA with the spindle in mouse oocytes

Project description:The oocytes of many species, both invertebrate and vertebrate, contain a large collection of localized determinants in the form of proteins and translationally inactive maternal mRNAs. However, it is unknown whether mouse oocytes contain localized MmRNA determinants and what mechanisms might be responsible for their control. We collected intact MII oocytes, enucleated MII oocyte cytoplasts (with the spindle removed), and spindle-chromosome complexes which had been microsurgically removed. RNA was extracted, amplified, labeled, and applied to microarrays to determine if any MmRNA determinants were localized to the SCC. We used microarrays to determine whether maternal mRNAs in mouse oocytes are enriched in the meiotic spindle

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Analysis of microRNAs in bovine early embryonic development

Project description:In animals, the maternal-to-embryonic transition (MET) is an important step occurring in the first days of early development. This important transition involves the degradation of maternal transcripts that have been stocked during oogenesis and used until this transition. Moreover, some precise and specific control mechanisms must govern the adequate synchronization of the MET events to promote embryonic genome activation. These mechanisms are not well understood, but microRNAs could be one of the mechanisms involved. MicroRNAs are short non-coding RNAs involved in a growing number of pathways by their post-transcriptional repression of mRNA translation. Thus, we analyzed microRNA expression during oocyte maturation by comparing GV-oocytes with MII-oocytes in cow. Groups of 300 Oocytes_GV and Oocytes_MII were used for miRNA expression analysis. The miRNA expressions were determined using the MiRMammalia_11.0 array from LC Sciences.

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Project description:Oocyte maturation is accompanied by a transition from mRNA stability to instability. We investigated the role of DCP1A and DCP2, proteins responsible for mRNA decapping, in mRNA destabilization during mouse oocyte maturation. siRNA-mediated knockdown of both Dcp1a and Dcp2 transcripts prior to initiation of maturation inhibited the maturation-associated increase of DCP1A and DCP2, stabilized a set of maternal mRNAs that are normally degraded during maturation, and inhibited development beyond the 2-cell stage, likely a consequence of failure to activate fully the zygotic genome. Total RNA from 30 MII eggs was used in each sample. Three independent biological replicates were analyzed for each condition.

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Quantitative proteomic analysis of oocytes from high-fat diet (HFD)-fed and normal diet mice

Project description:In order to establish an obese mouse model, female mice were continuously fed with a high-fat diet (HFD) or a normal diet (control) for 16 weeks beginning at three weeks of age. In this paper, these mice are termed ‘HFD mice’ and ‘control mice’, respectively. Accordingly, we call their oocytes ‘HFD oocytes’ and ‘control oocytes’. Substantial evidence indicates that the effects of maternal obesity on embryo/offspring development can be attributed to factors within the oocyte (9). To identify such potential effectors, we performed a comparative proteomic analysis of ovulated MII oocytes from control and HFD mice.

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Profiling and functional characterization of maternal mRNA translation in mouse maternal-to-zygotic transition

Project description:Translational regulation plays an important role in gene expression and function. Although the transcriptional dynamics of mouse pre-implantation embryos has been well characterized, the global mRNA translation landscape and the master regulators of zygotic genome activation (ZGA) remain unknown. Here, by developing and applying a low-input ribosome profiling (LiRibo-seq) technique, we profiled the mRNA translation landscape in mouse pre-implantation embryos and revealed the translational dynamics during mouse pre-implantation development. We identified a dramatic translational transition from MII oocytes to zygotes, and demonstrated that active translation of maternal mRNAs is essential for maternal-to-zygotic transition (MZT). We further showed that two maternal factors, Smarcd2 and Cyclin T2, whose translation is activated in zygotes, are required for chromatin reprogramming and ZGA, respectively. Our study thus not only filled in a knowledge gap on translational regulation during mammalian pre-implantation development, but also revealed new insights into the critical function of maternal mRNA translation in MZT.

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Comparison of the translational program in oocytes from wild type and Areg KO mice

Project description:The study tests the hypothesis that maternal mRNA translation in oocytes is sensitive to the environment in which the oocytes mature. Amphiregulin (AREG) is a critical signal for oocyte maturation but also for oocyte developmental competence. Here we have used a genome-wide approach to determine whether the oocyte translational program is affected when oocytes mature in vivo in the absence of AREG. To this aim, polysome arrays were used to define patterns of transcript recruitment to the polysomes in oocytes derived from wild type mice and mice homozygous null for the Areg gene. Forty-eight hours (h) after PMSG injection, mice were stimulated with hCG for 0, or 14 h, and GV, and MII stage oocytes were collected. Polysome bound mRNAs were purified, reverse-transcribed and linearly amplified with WT-Ovation FFPE RNA Amplification System V2 (NuGEN). 5µg cDNA were fragmented and hybridized with Affymetrix Mouse Genome 430.2 array chips. Experiments were done using 3 independent sample sets.

2013-10-02 | E-GEOD-46640 | biostudies-arrayexpress

RNA-seq of pabpn1l ko and wt female mice oocytes and zygotes

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2022-08-01 | E-MTAB-9488 | biostudies-arrayexpress

Effects of in vitro maturation on oocyte gene expression

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