Protein source in maturation media affects gene expression in cumulus cells and embryo development in cattle

2021 ◽  
pp. 1-14
Author(s):  
Nayara Ribeiro Kussano ◽  
Ligiane de Oliveira Leme ◽  
Margot Alves Nunes Dode
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Cumulus Cells ◽  
Protein Source

Effect of butafosfan supplementation during oocyte maturation on bovine embryo development

Zygote ◽  
2019 ◽  
Vol 27 (05) ◽  
pp. 321-328
Author(s):  
Lucas Teixeira Hax ◽  
Joao Alveiro Alvarado Rincón ◽  
Augusto Schneider ◽  
Lígia Margareth Cantarelli Pegoraro ◽  
Letícia Franco Collares ◽  
...  
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
Bovine Embryo ◽  
Cleavage Rate ◽  
Nuclear Maturation

SummaryAround 60–80% of oocytes maturated in vivo reached competence, while the proportion of maturation in vitro is rarely higher than 40%. In this sense, butafosfan has been used in vivo to improve metabolic condition of postpartum cows, and can represent an alternative to increase reproductive efficiency in cows. The aim of this study was to evaluate the addition of increasing doses of butafosfan during oocyte maturation in vitro on the initial embryo development in cattle. In total, 1400 cumulus–oocyte complexes (COCs) were distributed in four groups and maturated according to supplementation with increasing concentrations of butafosfan (0 mg/ml, 0.05 mg/ml, 0.1 mg/ml and 0.2 mg/ml). Then, 20 oocytes per group were collected to evaluate nuclear maturation and gene expression on cumulus cells and oocytes and the remaining oocytes were inseminated and cultured until day 7, when blastocysts were collected for gene expression analysis. A dose-dependent effect of butafosfan was observed, with decrease of cleavage rate and embryo development with higher doses. No difference between groups was observed in maturation rate and expression of genes related to oocyte quality. Our results suggest that butafosfan is prejudicial for oocytes, compromising cleavage and embryo development.

scholarly journals Is there a correlation between follicle size and gene expression in cumulus cells and is gene expression an indicator of embryo development?

2018 ◽  
Vol 16 (1) ◽  
Author(s):  
Semra Kahraman ◽  
Caroline Pirkevi Çetinkaya ◽  
Murat Çetinkaya ◽  
Mehmet Ali Tüfekçi ◽  
Cumhur Gökhan Ekmekçi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Cumulus Cells ◽  
Follicle Size

192 9-cis-RETINOIC ACID AFFECTS OOCYTE COMPETENCE MARKER GENE EXPRESSION IN THE BOVINE IN VITRO-MATURED OOCYTES AND CUMULUS CELLS

2012 ◽  
Vol 24 (1) ◽  
pp. 208
Author(s):  
G. K. Deb ◽  
S. R. Dey ◽  
J. I. Bang ◽  
I. K. Kong
Keyword(s):  
Gene Expression ◽  
Retinoic Acid ◽  
Embryo Development ◽  
Reference Genes ◽  
Cumulus Cells ◽  
Receptor Expression ◽  
Marker Genes ◽  
Oocyte Competence ◽  
Maturation Medium

Oocyte competence is the ability of an oocyte to undergo pre- and post-implantation development and to deliver a healthy offspring. A close association between the oocyte and the cumulus cells (CC) affects oocyte competence. Expression of several genes in the CC, known as oocyte competence markers, correlates with subsequent embryo development and quality. Addition of 9-cis-retinoic acid (9cisRA) to maturation medium increases oocyte competence through multiple mechanisms, including FSH/LH receptor expression, polyadenylation, growth factors signaling, oxidative-stress protection, or decreasing oocyte TNF-α gene expression. However, the effect of 9cisRA on the expression of oocyte competence markers in the oocytes and CC has not been determined. Therefore, the present study evaluated the effect of 9cisRA on the expression of oocyte competence marker genes in the oocytes and in the CC. Bovine cumulus–oocyte complexes, isolated from ovaries collected at the abattoir, were matured in vitro in the presence of 0 or 5 nM 9cisRA in the maturation medium (TCM-199 supplemented with 10% fetal bovine serum, 1 μg mL–1 of β-oestradiol, 10 μg mL–1 of follicle stimulating hormone, 0.6 mM cystein and 0.2 mM Na-pyruvate). After maturation, expression of target transcripts was quantified in CC and zona-free oocytes by SYBER green real-time PCR. The expression was normalized against a minimum of 2 out of 4 reference genes analyzed each time with target genes. The best combination of reference genes was automatically calculated by the CFX manager V1.1 program (BioRad) based on M-value during the analysis of gene expression data. A minimum of 5 biological replicates (50–60 oocytes/replicate) were performed for statistical analysis using a Student's t-test. Results indicated that 9cisRA increased (P < 0.05 to P < 0.001) expression of gremlin 1, prostaglandin G/H synthase 2, pentraxin 3, midkine, CD9 and thioredoxin mRNA in oocytes (3.0-, 2.8-, 3.1-, 2.0-, 4.0- and 2.4-fold) and CC (4.0-, 3.2-, 1.9-, 1.7-, 4.0- and 2.4-fold) compared to controls. In contrast, the aldose reductase 1b1 mRNA was down-regulated both in oocytes (1.0- vs 2.0-fold; P < 0.05) and in CC (1.0- vs 1.9-fold; P < 0.04) compared to the respective controls. In conclusion, the present study indicates that 9cisRA influences mRNA expression of oocytes and cumulus cells. This might be another explanation of the improved embryo development and quality in response to 9cisRA during in vitro maturation. This work was partly supported by a scholarship from the BK21 program, the KRF (KRF-2008-211-F00011), the Next-generation BioGreen 21 Program (No. PJ007990012011), IPET (110020-3 and 109016-3) and the KOSEF (10525010001-05N2501-00110).

scholarly journals Fatty acid induced lipolysis influences embryo development, gene expression and lipid droplet formation in the porcine cumulus cells†

2020 ◽  
Vol 103 (1) ◽  
pp. 36-48
Author(s):  
Piotr Pawlak ◽  
Natalia Malyszka ◽  
Izabela Szczerbal ◽  
Pawel Kolodziejski
Keyword(s):  
Gene Expression ◽  
Fatty Acids ◽  
Oleic Acid ◽  
Fatty Acid ◽  
Stearic Acid ◽  
Embryo Development ◽  
Follicular Fluid ◽  
Lipid Droplets ◽  
Saturated Fatty Acids ◽  
Cumulus Cells

Abstract The pig oocyte maturation protocol differs from other mammalian species due to dependence on follicular fluid (FF) supplementation. One of the most abundant components of the porcine follicular fluid are fatty acids (FAs). Although evidence from other mammalian models revealed a negative impact of saturated fatty acids (SFA) on developmental competence of oocytes, pig has not yet been widely analyzed. Therefore, we aimed to investigate whether supplementation of IVM medium with 150 μM of stearic acid (SA) and oleic acid (OA) affects lipid content and expression of genes related to fatty acid metabolism in porcine cumulus–oocyte complexes and parthenogenetic embryo development. We found significant influence of fatty acids on lipid metabolism in cumulus cells without affecting the oocyte proper. The expression of ACACA, SCD, PLIN2, FADS1, and FADS2 genes was upregulated (P &lt; 0.01) in cumulus cells, while their expression in oocytes did not change. The increase in gene expression was more pronounced in the case of OA (e.g., up to 30-fold increase in PLIN2 transcript level compared to the control). The number of lipid droplets and occupied area increased significantly in the cumulus cells and did not change in oocytes after SA treatment. Oleic acid improved the blastocyst rate (48 vs 32% in control), whereas stearic acid did not affect this parameter (27%). Additionally, we have discovered a phenotypic diversity of LD in cumulus cells in response to FA supplementation, suggesting extensive lipolysis in response to SA. Stearic acid excess in maturation media led to the formation of multiple micro lipid droplets in cumulus cells.

scholarly journals Heat Shock Protein 70 Improves In Vitro Embryo Yield and Quality from Heat Stressed Bovine Oocytes

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1794
Author(s):  
Konstantina Stamperna ◽  
Themistoklis Giannoulis ◽  
Eleni Dovolou ◽  
Maria Kalemkeridou ◽  
Ioannis Nanas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Heat Shock Protein 70 ◽  
Cumulus Cells ◽  
Intramolecular Interactions ◽  
Developmental Competence ◽  
Embryo Yield ◽  
Bovine Oocytes

Heat shock protein 70 (HSP70) is a chaperon that stabilizes unfolded or partially folded proteins, preventing inappropriate inter- and intramolecular interactions. Here, we examined the developmental competence of in vitro matured oocytes exposed to heat stress with or without HSP70. Bovine oocytes were matured for 24 h at 39 °C without (group C39) or with HSP70 (group H39) and at 41 °C for the first 6 h, followed by 16 h at 39 °C with (group H41) or without HSP70 (group C41). After insemination, zygotes were cultured for 9 days at 39 °C. Cleavage and embryo yield were assessed 48 h post insemination and on days 7, 8, 9, respectively. Gene expression was assessed by RT-PCR in oocytes, cumulus cells and blastocysts. In C41, blastocysts formation rate was lower than in C39 and on day 9 it was lower than in H41. In oocytes, HSP70 enhanced the expression of three HSP genes regardless of incubation temperature. HSP70 at 39 °C led to tight coordination of gene expression in oocytes and blastocysts, but not in cumulus cells. Our results imply that HSP70, by preventing apoptosis, supporting signal transduction, and increasing antioxidant protection of the embryo, protects heat stressed maturing bovine oocyte and restores its developmental competence.

scholarly journals Ancient Sturgeons Possess Effective DNA Repair Mechanisms: Influence of Model Genotoxicants on Embryo Development of Sterlet, Acipenser ruthenus

2020 ◽  
Vol 22 (1) ◽  
pp. 6
Author(s):  
Ievgeniia Gazo ◽  
Roman Franěk ◽  
Radek Šindelka ◽  
Ievgen Lebeda ◽  
Sahana Shivaramu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Damage ◽  
Dna Repair ◽  
Embryo Development ◽  
Hatching Rate ◽  
Embryo Survival ◽  
Dna Repair Mechanisms ◽  
Acipenser Ruthenus ◽  
Repair Mechanisms ◽  
Sterlet Acipenser Ruthenus

DNA damage caused by exogenous or endogenous factors is a common challenge for developing fish embryos. DNA damage repair (DDR) pathways help organisms minimize adverse effects of DNA alterations. In terms of DNA repair mechanisms, sturgeons represent a particularly interesting model due to their exceptional genome plasticity. Sterlet (Acipenser ruthenus) is a relatively small species of sturgeon. The goal of this study was to assess the sensitivity of sterlet embryos to model genotoxicants (camptothecin, etoposide, and benzo[a]pyrene), and to assess DDR responses. We assessed the effects of genotoxicants on embryo survival, hatching rate, DNA fragmentation, gene expression, and phosphorylation of H2AX and ATM kinase. Exposure of sterlet embryos to 1 µM benzo[a]pyrene induced low levels of DNA damage accompanied by ATM phosphorylation and xpc gene expression. Conversely, 20 µM etoposide exposure induced DNA damage without activation of known DDR pathways. Effects of 10 nM camptothecin on embryo development were stage-specific, with early stages, before gastrulation, being most sensitive. Overall, this study provides foundational information for future investigation of sterlet DDR pathways.

Serial analysis of gene expression (SAGE) during porcine embryo development

2004 ◽  
Vol 16 (2) ◽  
pp. 87 ◽  
Author(s):  
Le Ann Blomberg ◽  
Kurt A. Zuelke
Keyword(s):  
Gene Expression ◽  
Functional Genomics ◽  
Embryo Development ◽  
Molecular Mechanisms ◽  
Physiological Role ◽  
Transgenic Animals ◽  
Specific Gene ◽  
Research Strategies

Functional genomics provides a powerful means for delving into the molecular mechanisms involved in pre-implantation development of porcine embryos. High rates of embryonic mortality (30%), following either natural mating or artificial insemination, emphasise the need to improve the efficiency of reproduction in the pig. The poor success rate of live offspring from in vitro-manipulated pig embryos also hampers efforts to generate transgenic animals for biotechnology applications. Previous analysis of differential gene expression has demonstrated stage-specific gene expression for in vivo-derived embryos and altered gene expression for in vitro-derived embryos. However, the methods used to date examine relatively few genes simultaneously and, thus, provide an incomplete glimpse of the physiological role of these genes during embryogenesis. The present review will focus on two aspects of applying functional genomics research strategies for analysing the expression of genes during elongation of pig embryos between gestational day (D) 11 and D12. First, we compare and contrast current methodologies that are being used for gene discovery and expression analysis during pig embryo development. Second, we establish a paradigm for applying serial analysis of gene expression as a functional genomics tool to obtain preliminary information essential for discovering the physiological mechanisms by which distinct embryonic phenotypes are derived.

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