Folliculogenesis and acquisition of oocyte competence in cows.

IVF success depends on hundreds of factors and details but the oocyte quality remains the most important and problematic issue. All antral follicles contain oocytes and all of them have that have reached their full size, can be aspirated, can mature and can be fertilized in vitro. But only a few will make it to embryo unless harvested at a very specific time/status. The conditions impacting the oocyte competence are essentially dependant on the follicular status. Growing follicles contains oocytes that have not completed their preparation, as they are still writing information (RNA), later, dominant follicles or follicles at the plateau phase, stop transcription and become candidates for development. Once in transcriptional arrest, the oocytes, if not ovulated in a short amount of time, do not always make good embryos. This window is affected by time and follicle size and looks like a bell curve. The following review further explain the physiological and molecular evidences that we have to illustrate the competence window and provides clues on how to optimize ovarian stimulation to maximise oocyte quality.

Folliculogenesis and acquisition of oocyte competence in cows

IVF success depends on hundreds of factors and details but the oocyte quality remains the most important and problematic issue. All antral follicles contain oocytes and all of them have that have reached their full size, can be aspirated, can mature and can be fertilized in vitro. But only a few will make it to embryo unless harvested at a very specific time/status. The conditions impacting the oocyte competence are essentially dependant on the follicular status. Growing follicles contains oocytes that have not completed their preparation, as they are still writing information (RNA), later, dominant follicles or follicles at the plateau phase, stop transcription and become candidates for development. Once in transcriptional arrest, the oocytes, if not ovulated in a short amount of time, do not always make good embryos. This window is affected by time and follicle size and looks like a bell curve. The following review further explain the physiological and molecular evidences that we have to illustrate the competence window and provides clues on how to optimize ovarian stimulation to maximise oocyte quality.

Folliculogenesis and acquisition of oocyte competence in cows

IVF success depends on hundreds of factors and details but the oocyte quality remains the most important and problematic issue. All antral follicles contain oocytes and all of them have that have reached their full size, can be aspirated, can mature and can be fertilized in vitro. But only a few will make it to embryo unless harvested at a very specific time/status. The conditions impacting the oocyte competence are essentially dependant on the follicular status. Growing follicles contains oocytes that have not completed their preparation, as they are still writing information (RNA), later, dominant follicles or follicles at the plateau phase, stop transcription and become candidates for development. Once in transcriptional arrest, the oocytes, if not ovulated in a short amount of time, do not always make good embryos. This window is affected by time and follicle size and looks like a bell curve. The following review further explain the physiological and molecular evidences that we have to illustrate the competence window and provides clues on how to optimize ovarian stimulation to maximise oocyte quality.(AU)

Genome-wide screening of DNA methylation in bovine blastocysts with different kinetics of development.

BACKGROUND: The timing of the first cell divisions may predict the developmental potential of an embryo, including its ability to establish pregnancy. Besides differences related to metabolism, stress, and survival, embryos with different speeds of development present distinct patterns of gene expression, mainly related to energy and lipid metabolism. As gene expression is regulated by epigenetic factors, and that includes DNA methylation patterns, in this study we compared the global DNA methylation profile of embryos with different kinetics of development in order to identify general pathways and regions that are most influenced by this phenotype. For this purpose, bovine embryos were in vitro produced using sexed semen (female), classified as fast (four or more cells) or slow (two cells) at 40 hpi and cultured until blastocyst stage, when they were analyzed. RESULTS: Genome-wide DNA methylation analysis identified 11,584 differently methylated regions (DMRs) (7976 hypermethylated regions in fast and 3608 hypermethylated regions in slow embryos). Fast embryos presented more regions classified as hypermethylated distributed throughout the genome, as in introns, exons, promoters, and repeat elements while in slow embryos, hypermethylated regions were more present in CpG islands. DMRs were clustered by means of biological processes, and the most affected pathways were related to cell survival/differentiation and energy/lipid metabolism. Transcripts profiles from DM genes connected with these pathways were also assessed, and the most part disclosed changes in relative quantitation. CONCLUSION: The kinetics of the first cleavages influences the DNA methylation and expression profiles of genes related to metabolism and differentiation pathways and may affect embryo viability.

Effect of vitrification using the Cryotop method on the gene expression profile of in vitro-produced bovine embryos.

The present study analyzed the changes in gene expression induced by the Cryotop vitrification technique in bovine blastocyst-stage embryos, using Agilent EmbryoGENE microarray slides. Bovine in vitro-produced embryos were vitrified and compared with nonvitrified (control) embryos. After vitrification, embryos were warmed and cultured for an additional 4 hours. Survived embryos were used for microarray analysis and quantitative polymerase chain reaction (qPCR) quantification. Survival rates were higher (P < 0.05) in the control embryos (100%) than in the vitrified embryos (87%). Global gene expression analysis revealed that only 43 out of 21,139 genes exhibited significantly altered expression in the vitrified embryos compared to the control embryos, with a very limited fold change (P < 0.05). A total of 10 genes were assessed by qPCR. Only the FOS-like antigen 1 (FOSL1) gene presented differential expression (P < 0.05) according to both the array and qPCR methods, and it was overexpressed in vitrified embryos. Although, the major consequence of vitrification seems to be the activation of the apoptosis pathway in some cells. Indeed, FOSL1 is part of the activating protein 1 transcription factor complex and is implicated in a variety of cellular processes, including proliferation, differentiation, and apoptosis. Therefore, our results suggest that a limited increase in the rate of apoptosis was the only detectable response of the embryos to vitrification stress.

Quantitative expression of candidate genes for developmental competence in bovine two-cell embryos.

Only competent oocytes are able to undergo complete maturation and normal embryonic development. Therefore, the identification of genes that are differentially expressed in competent oocytes would contribute to our understanding of the factors controlling competency. It is well known that time of cleavage after insemination in vitro is highly correlated with embryonic developmental potential and this can be used to distinguish between oocytes of different quality. The main objective of this study was to identify genes associated with competency and rapid cleavage. We examined the expression of 16 candidate genes (IDH, YEAF Cathepsin B, RAD50, TCP1 NCOR1, HUEL, STK6, ZNF403, AOP2, EEF1A1, Hsp90, Hsp40, AKR1B1, PGRMC1, and DMRT2) in early and late cleaving embryos, by real time PCR. These transcripts were derived from previous study in our laboratory using cDNA coming from a suppressive subtraction hybridization (SSH) between early cleaving versus late cleaving embryos spotted on a microarray slide. Of the 16 genes evaluated, 3 (IDH, YEAF, and H2A) showed statistical difference (P < 0.05) between early and late cleaving embryos. However, some genes such as Cathepsin B (P = 0.0677), RAD50 (P = 0.0899), and TCP1 (P = 0.0824) tended to show higher expression in the early cleaving than in the late cleaving embryo. In conclusion, we have identified three genes (YEAF, IDH, H2A) that were differentially expressed in the early cleaving embryos, and their expression can be associated with greater developmental competence.