Embryonic poly(A)‐binding protein is differently expressed and interacts with the messenger RNAs in the mouse oocytes and early embryos

2018 ◽  
Vol 120 (3) ◽  
pp. 4694-4709 ◽  
Author(s):  
Fatma Uysal ◽  
Saffet Ozturk
Keyword(s):  
Binding Protein ◽  
Messenger Rnas ◽  
Mouse Oocytes ◽  
Early Embryos

Gene expression during mammalian oogenesis and early embryogenesis: quantification of three messenger RNAs abundant in fully grown mouse oocytes

Development ◽  
1989 ◽  
Vol 106 (2) ◽  
pp. 251-261 ◽  
Author(s):  
R.J. Roller ◽  
R.A. Kinloch ◽  
B.Y. Hiraoka ◽  
S.S. Li ◽  
P.M. Wassarman
Keyword(s):  
Gene Expression ◽  
Steady State ◽  
Early Embryogenesis ◽  
Messenger Rnas ◽  
Oocyte Growth ◽  
Mouse Oocytes ◽  
Mouse Tissues ◽  
Specific Mrnas ◽  
Steady State Levels ◽  
Ribonuclease Protection

Ribonuclease protection assays have been used to quantitatively assess changes in steady-state levels of specific mRNAs during oogenesis and early embryogenesis in mice. The mRNAs encode ZP3 (a glycoprotein that serves as a sperm receptor), LDH-B (heart-type lactate dehydrogenase), and MOM-1 (a protein of unknown function). MOM-1 and LDH-B are expressed in a variety of adult mouse tissues and midgestation embryos, whereas ZP3 expression is restricted completely to oocytes. All three mRNAs are expressed by growing mouse oocytes and accumulate to unusually high levels in fully grown oocytes as compared to somatic cells; 240,000, 200,000 and 74,000 copies mRNA per fully grown oocyte for ZP3, LDH-B and MOM-1, respectively. Steady-state levels of LDH-B and MOM-1 mRNA undergo a modest decline (approximately 20–40%) during ovulation when fully grown oocytes become unfertilized eggs and, in general, mirror the reported change in poly(A)+RNA levels during this period of development. On the other hand, the level of ZP3 mRNA declines dramatically (approximately 98%) during ovulation, from approximately 240,000 copies per oocyte to approximately 5000 copies per unfertilized egg, and ZP3 mRNA is undetectable in fertilized eggs (less than 1000 copies per fertilized egg). MOM-1 mRNA is expressed at relatively low levels in morulae (approximately 2000 copies per embryo) and blastocysts (approximately 5000 copies per embryo), whereas ZP3 mRNA remains undetectable (less than 1000 copies per embryo) at these stages of preimplantation development. These findings are discussed in the context of overall gene expression during oocyte growth, meiotic maturation and early embryogenesis in mice.

A monoclonal antibody against mouse oocyte cytoskeleton recognizing cytokeratin-type filaments

Development ◽  
1985 ◽  
Vol 90 (1) ◽  
pp. 197-209
Author(s):  
E. Lehtonen
Keyword(s):  
Epithelial Cells ◽  
Immunofluorescence Microscopy ◽  
Staining Pattern ◽  
Blastocyst Stage ◽  
Tissue Sections ◽  
Mouse Oocytes ◽  
Thick Filaments ◽  
Trophectoderm Cells ◽  
Early Embryos ◽  
Cytokeratin Filaments

Monoclonal antibodies were raised to detergent-extracted cytoskeleton preparations of mouse oocytes. In immunofluorescence microscopy, one of the antibodies, OCS-1, localizes exclusively to epithelial cells in frozen tissue sections, including various simple and stratified epithelia. The antibody decorates a keratin-type of fibrillax, vinblastine-resistant network in various cultured, epithelial-type cells, but not in myoid or fibroblastoid cells. In mouse oocytes and cleavage-stage embryos, the OCS-1 antibody gives a diffuse, spotty staining pattern. In blastocyst-stage embryos, the antibody reveals a keratin-type filamentous organization in the trophectoderm cells. In immunoelectron microscopy, the OCS-1 antibody decorates 10 nm-thick filaments, often identifiable as desmosome-attached tonofilaments, in detergent-treated trophectoderm cells. The antigen(s) recognized by the OCS-1 antibody is apparently present in, or closely associated with, cytokeratin filaments. In addition to mouse oocytes and early embryos, a wide variety of epithelial cells in various species seem to share this antigen(s). The present results suggest that at the early stages, the cytokeratin-related antigen(s) defined by the OCS-1 antibody are stored in a non-fibrillar form which is then converted into a fibrillar network at the blastocyst stage. A pre-existing supply of cytokeratin-related protein may be essential for the development of the blastocyst.

scholarly journals Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ

2020 ◽  
Vol 48 (8) ◽  
pp. 4507-4520 ◽  
Author(s):  
Smriti Pandey ◽  
Chandra M Gravel ◽  
Oliver M Stockert ◽  
Clara D Wang ◽  
Courtney L Hegner ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Molecular Mechanisms ◽  
Rna Binding ◽  
Binding Protein ◽  
Rna Binding Protein ◽  
Site Directed Mutagenesis ◽  
Genetic Identification ◽  
Messenger Rnas ◽  
Functional Surface

Abstract The FinO-domain-protein ProQ is an RNA-binding protein that has been known to play a role in osmoregulation in proteobacteria. Recently, ProQ has been shown to act as a global RNA-binding protein in Salmonella and Escherichia coli, binding to dozens of small RNAs (sRNAs) and messenger RNAs (mRNAs) to regulate mRNA-expression levels through interactions with both 5′ and 3′ untranslated regions (UTRs). Despite excitement around ProQ as a novel global RNA-binding protein, and its potential to serve as a matchmaking RNA chaperone, significant gaps remain in our understanding of the molecular mechanisms ProQ uses to interact with RNA. In order to apply the tools of molecular genetics to this question, we have adapted a bacterial three-hybrid (B3H) assay to detect ProQ’s interactions with target RNAs. Using domain truncations, site-directed mutagenesis and an unbiased forward genetic screen, we have identified a group of highly conserved residues on ProQ’s NTD as the primary face for in vivo recognition of two RNAs, and propose that the NTD structure serves as an electrostatic scaffold to recognize the shape of an RNA duplex.

RNAi Experiments in Mouse Oocytes and Early Embryos

2009 ◽  
Vol 2009 (1) ◽  
pp. pdb.top56-pdb.top56 ◽  
Author(s):  
P. Svoboda ◽  
P. Stein
Keyword(s):  
Mouse Oocytes ◽  
Early Embryos

scholarly journals Embryonic Poly(A)-Binding Protein Stimulates Translation in Germ Cells

2005 ◽  
Vol 25 (5) ◽  
pp. 2060-2071 ◽  
Author(s):  
Gavin S. Wilkie ◽  
Philippe Gautier ◽  
Diane Lawson ◽  
Nicola K. Gray
Keyword(s):  
Xenopus Laevis ◽  
Oocyte Maturation ◽  
Germ Cells ◽  
Binding Protein ◽  
Expression Patterns ◽  
General Role ◽  
Early Embryos ◽  
Relative Divergence

ABSTRACT The function of poly(A)-binding protein 1 (PABP1) in poly(A)-mediated translation has been extensively characterized. Recently, Xenopus laevis oocytes and early embryos were shown to contain a novel poly(A)-binding protein, ePABP, which has not been described in other organisms. ePABP was identified as a protein that binds AU-rich sequences and prevents shortening of poly(A) tails. Here, we show that ePABP is also expressed in X. laevis testis, suggesting a more general role for ePABP in gametogenesis. We find that ePABP is conserved throughout vertebrates and that mouse and X. laevis cells have similar tissue-specific ePABP expression patterns. Furthermore, we directly assess the role of ePABP in translation. We show that ePABP is associated with polysomes and can activate the translation of reporter mRNAs in vivo. Despite its relative divergence from PABP1, we find that ePABP has similar functional domains and can bind to several PABP1 partners, suggesting that they may use similar mechanisms to activate translation. In addition, we find that PABP1 and ePABP can interact, suggesting that these proteins may be bound simultaneously to the same mRNA. Finally, we show that the activity of both PABP1 and ePABP increases during oocyte maturation, when many mRNAs undergo polyadenylation.

scholarly journals Why mouse oocytes and early embryos ignore miRNAs?

RNA Biology ◽  
2010 ◽  
Vol 7 (5) ◽  
pp. 559-563 ◽  
Author(s):  
Petr Svoboda
Keyword(s):  
Mouse Oocytes ◽  
Early Embryos

DNA methylation pattern in mouse oocytes and their in vitro fertilized early embryos: effect of oocyte vitrification

Zygote ◽  
2012 ◽  
Vol 22 (2) ◽  
pp. 138-145 ◽  
Author(s):  
Ying Liang ◽  
Xiang-Wei Fu ◽  
Jun-Jie Li ◽  
Dian-Shuai Yuan ◽  
Shi-En Zhu
Keyword(s):  
Dna Methylation ◽  
Fluorescein Isothiocyanate ◽  
Methylation Pattern ◽  
Oocyte Vitrification ◽  
Developmental Potential ◽  
Mouse Oocytes ◽  
Early Embryos ◽  
Vitrification Solution ◽  
Early Mouse

SummaryThis study was conducted to investigate the pattern of DNA methylation in vitrified–thawed mouse oocytes and their in vitro fertilized early embryos. Firstly, mouse oocytes at metaphase II (MII) stage of meiosis were allocated randomly into three groups: (1) untreated (control); (2) exposed to vitrification solution without being plunged into liquid nitrogen (toxicity); or (3) vitrified by open-pulled straw (OPS) method (vitrification). Oocytes from all three groups were fertilized subsequently in vitro. The level of DNA methylation in the MII oocytes and their early embryos was then examined by immunofluorescence using an anti-5-methylcytosine (anti-5-MeC) monoclonal antibody and fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Developmental rates to 2-cell embryos (62.28%) and blastocysts (43.68%) of the vitrified–thawed oocytes were lower (P < 0.01) than those of fresh oocytes (81.47%, 61.99%) and vitrification solution treated (79.20%, 60.04%) oocytes. DNA methylation (as reflected by 5-MeC fluorescence intensity) in the vitrification group was less (P < 0.01) for MII oocyte and 2- to 8-cell stages compared with that in the control and toxicity groups. Accordingly, a reduction in global genomic methylation due to vitrification of MII oocytes may result in compromised in vitro developmental potential in early mouse embryos.

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