Messenger RNA expression of Pabpnl1 and Mbd3l2 genes in oocytes and cleavage embryos

Biase, Fernando H.; Martelli, Lúcia Regina; Puga, Renato; Giuliatti, Silvana; Santos-Biase, Weruska Karyna Freitas; Merighe, Giovana Krempel Fonseca; Meirelles, Flávio Vieira

doi:10.1016/j.fertnstert.2009.08.051

Cited by 7 publications

(7 citation statements)

References 39 publications

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“…COCs were then selected from the follicular fluid and washed in TCM-199 medium supplemented with 0.42 M sodium bicarbonate (Macron), 0.02 M Hepes (Sigma-Aldrich), 10% (v:v) fetal bovine serum (Seradigm), 0.05 g/ml gentamicin (Amresco), 0.022 g/ml pyruvate (Acros Organics) and 1× Glutamax (Gibco). COCs were selected based on morphological characteristics indicative of a greater developmental potential [ 21 , 37 ]. The obtained COCs containing oocytes with homogeneous cytoplasm surrounded by more than five layers of cumulus cells were used in our research.…”

Section: Methodsmentioning

confidence: 99%

Functional signaling and gene regulatory networks between the oocyte and the surrounding cumulus cells

Biase

Kimble

2018

BMC Genomics

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BackgroundThe maturation and successful acquisition of developmental competence by an oocyte, the female gamete, during folliculogenesis is highly dependent on molecular interactions with somatic cells. Most of the cellular interactions identified, thus far, are modulated by growth factors, ions or metabolites. We hypothesized that this interaction is also modulated at the transcriptional level, which leads to the formation of gene regulatory networks between the oocyte and cumulus cells. We tested this hypothesis by analyzing transcriptome data from single oocytes and the surrounding cumulus cells collected from antral follicles employing an analytical framework to determine interdependencies at the transcript level.ResultsWe overlapped our transcriptome data with putative protein-protein interactions and identified hundreds of ligand-receptor pairs that can transduce paracrine signaling between an oocyte and cumulus cells. We determined that 499 ligand-encoding genes expressed in oocytes and cumulus cells are functionally associated with transcription regulation (FDR < 0.05). Ligand-encoding genes with specific expression in oocytes or cumulus cells were enriched for biological functions that are likely associated with the coordinated formation of transzonal projections from cumulus cells that reach the oocyte’s membrane. Thousands of gene pairs exhibit significant linear co-expression (absolute correlation > 0.85, FDR < 1.8 × 10− 5) patterns between oocytes and cumulus cells. Hundreds of co-expressing genes showed clustering patterns associated with biological functions (FDR < 0.5) necessary for a coordinated function between the oocyte and cumulus cells during folliculogenesis (i.e. regulation of transcription, translation, apoptosis, cell differentiation and transport).ConclusionOur analyses revealed a complex and functional gene regulatory circuit between the oocyte and surrounding cumulus cells. The regulatory profile of each cumulus-oocyte complex is likely associated with the oocytes’ developmental potential to derive an embryo.Electronic supplementary materialThe online version of this article (10.1186/s12864-018-4738-2) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Functional signaling and gene regulatory networks between the oocyte and the surrounding cumulus cells

Biase

Kimble

2018

BMC Genomics

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“…The molecular mechanisms that govern oocyte competence are mostly still unknown. However, some oocyte-specific genes have been described revealing their importance in promoting embryogenesis (Katz-Jaffe et al , 2009; Biase et al , 2010; Belli et al , 2013; Bessa et al , 2013; Biase et al , 2014). The profiling of gene expression in oocytes during maturation may help us understand the regulation of oocyte competence to mature and to sustain embryo development during the first two cleavages (Fair et al , 2007).…”

Section: Introductionmentioning

confidence: 99%

In vitromaturation alters gene expression in bovine oocytes

Adona

Leal

Biase

et al. 2016

Zygote

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Gene expression profiling of in vivo- and in vitro-matured bovine oocytes can identify transcripts related to the developmental potential of oocytes. Nonetheless, the effects of in vitro culturing oocytes are yet to be fully understood. We tested the effects of in vitro maturation on the transcript profile of oocytes collected from Bos taurus indicus cows. We quantified the expression of 1488 genes in in vivo- and in vitro-matured oocytes. Of these, 51 genes were up-regulated, whereas 56 were down-regulated (≥2-fold) in in vivo-matured oocytes in comparison with in vitro-matured oocytes. Quantitative real-time polymerase chain reaction (PCR) of nine genes confirmed the microarray results of differential expression between in vivo- and in vitro-matured oocytes (EZR, EPN1, PSEN2, FST, IGFBP3, RBBP4, STAT3, FDPS and IRS1). We interrogated the results for enrichment of Gene Ontology categories and overlap with protein-protein interactions. The results revealed that the genes altered by in vitro maturation are mostly related to the regulation of oocyte metabolism. Additionally, analysis of protein-protein interactions uncovered two regulatory networks affected by the in vitro culture system. We propose that the differentially expressed genes are candidates for biomarkers of oocyte competence. In vitro oocyte maturation can affect the abundance of specific transcripts and are likely to deplete the developmental competence.

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“…The abundance of specific ribonucleic acids (RNAs) stored in the female gamete, namely the oocyte, has a direct relationship with the acquisition of developmental competence [ 1 – 3 ]. With the recent advances of next generation sequencing, single-oocyte RNA sequencing data have been generated for mice [ 4 – 6 ], cattle [ 7 ], goats [ 8 ], and humans [ 9 ].…”

Section: Introductionmentioning

confidence: 99%

Extraction of total RNA from single-oocytes and single-cell mRNA sequencing of swine oocytes

2018

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ObjectiveAnalyses of single oocytes are essential for a fine dissection of molecular features governing developmental competence. We adapted the phenol–chloroform procedure for the purification of total RNA from single oocytes.ResultsKey modifications include the use of Phasemaker™ tubes, a second chloroform wash of the aqueous phase, and the precipitation of the RNA with glyclogen in a 200 μl micro-centrifuge tube. Assessment of the RNA profile from single oocytes showed distinct peaks for 18S and 28S ribosomal subunits. This approach permitted the extraction of small RNAs from single oocytes, which was evident by the presence of 5S and 5.8S rRNAs and tRNAs around 122–123 nucleotides long. The amplification of polyadenylated RNA resulted in detectable DNA products ranging from ~ 500 to ~ 5000 nucleotides. We used the amplified DNA as template for single-cell mRNA-sequencing of five swine oocytes and quantified the expression levels of 9587 genes with complete coverage of transcripts over 10,000 nucleotides in length. The coverage was similar in all oocytes sequenced, demonstrating consistent high RNA quality across samples. We isolated total RNA from single oocytes and demonstrated that the quality was appropriate for single-cell mRNA-sequencing.Electronic supplementary materialThe online version of this article (10.1186/s13104-018-3264-2) contains supplementary material, which is available to authorized users.

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Messenger RNA expression of Pabpnl1 and Mbd3l2 genes in oocytes and cleavage embryos

Cited by 7 publications

References 39 publications

Functional signaling and gene regulatory networks between the oocyte and the surrounding cumulus cells

Functional signaling and gene regulatory networks between the oocyte and the surrounding cumulus cells

In vitromaturation alters gene expression in bovine oocytes

Extraction of total RNA from single-oocytes and single-cell mRNA sequencing of swine oocytes

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