Gene Expression Analysis of Bovine Oocytes With High Developmental Competence Obtained From FSH-Stimulated Animals

2013 ◽  
pp. n/a-n/a ◽  
Author(s):  
RÉMI LABRECQUE ◽  
CHRISTIAN VIGNEAULT ◽  
PATRICK BLONDIN ◽  
MARC-ANDRÉ SIRARD
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Developmental Competence ◽  
Bovine Oocytes

Gene Expression Analysis of Bovine Oocytes with High Developmental Competence in Super-Stimulation Protocol.

2011 ◽  
Vol 85 (Suppl_1) ◽  
pp. 331-331
Author(s):  
Remi Labrecque ◽  
Audrey Bunel ◽  
Anne-Laure Nivet ◽  
Josee Belanger ◽  
Christian Vigneault ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Developmental Competence ◽  
Stimulation Protocol ◽  
Bovine Oocytes

Gene expression analysis of bovine oocytes at optimal coasting time combined with GnRH antagonist during the no-FSH period

2014 ◽  
Vol 81 (8) ◽  
pp. 1092-1100 ◽  
Author(s):  
Rémi Labrecque ◽  
Christian Vigneault ◽  
Patrick Blondin ◽  
Marc-André Sirard
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Gnrh Antagonist ◽  
Bovine Oocytes

307 MATURATION RATE AND GENE EXPRESSION ANALYSIS OF GOAT OOCYTES SELECTED BY FOLLICLE SIZE AND BRILLIANT CRESYL BLUE STAINING

2015 ◽  
Vol 27 (1) ◽  
pp. 242
Author(s):  
M. Yang ◽  
S. Hu ◽  
L. Cox ◽  
M. Regouski ◽  
H. Rutigliano ◽  
...  
Keyword(s):  
Gene Expression ◽  
Oocyte Maturation ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Oocyte Quality ◽  
Developmental Competence ◽  
Brilliant Cresyl Blue ◽  
Relative Expression ◽  
Cresyl Blue ◽  
Maturation Rate

Oocyte quality plays a critical role in determining the success of embryo development. Studies on cattle and goats indicate that oocytes derived from large follicles (LFO) have greater developmental competence than those derived from small follicles (SFO). Brilliant cresyl blue (BCB) staining determines the activity of glucose-6-phosphate dehydrogenase and is a commonly used noninvasive marker of oocyte competence. Studies in pigs, goats, cows, mice, and dogs showed that the maturation and blastocyst developmental rate of BCB+ oocytes is significantly higher than BCB– oocytes. The aim of this study was to evaluate the maturation rate of goat oocytes selected based on follicular size and BCB staining and compare their relative patterns of gene expression. Maturation rate and gene expression profile were expected to be different in these oocyte groups. Cumulus-oocyte complexes were recovered from abattoir-derived ovaries using a slicing technique. Eleven rounds of oocyte maturation and 4 rounds of BCB staining were carried out. During each replicate, oocytes from large (≥3 mm) and small (<3 mm) follicles were collected separately from the same group of ovaries. Oocyte maturation rates were 54.3 ± 5.4% for LFO (n = 378) and only 33.5 ± 3.7% for SFO (n = 981; P < 0.01). The BCB+ (n = 223) oocytes yielded a significantly higher maturation rate than the BCB– (n = 194) oocytes (56.1 ± 1.8 v. 20.6 ± 3.8%, respectively; P < 0.001). Gene expression analysis was conducted on individual MII oocytes (21 oocytes per group). Specific target amplification was performed on a single oocyte directly by using the CellsDirect One-Step qRT–PCR Kit (Invitrogen). Quantitative real-time PCR was then performed using the 48.48 BioMark platform from Fluidigm. Forty two genes were selected from the following categories: growth factors, transcription factors, metabolism, pluripotency, cell cycle, apoptosis, and oocyte-specific genes. Relative expression values were calculated using the ΔΔCT (fold change) method and analysed by ANOVA. The significance was assigned at P < 0.05. The relative expression of CCNA2, CDK2, CCNB1, POU5F1, SOX2, EGF, FGF2, GDF9, ZP3, BCL2, GJA1, DDR1, PFKFB3, IGF2R, and GRB10 was significantly greater (P < 0.05) in both LFO and BCB+ oocytes compared to SFO and BCB– oocytes, respectively. The proapoptotic gene BAX, the ACSL3 gene involved in fatty acid oxidation, and the growth factor IGF1 were expressed significantly higher (P < 0.05) in SFO compared to LFO. By investigating these differentially expressed transcripts, we will better understand pathways involved in oocyte developmental competence and potentially use them as markers of oocyte quality. We expect that the ability to select oocytes of better quality based on BCB staining will improve outcomes of IVF and SCNT.

Gene expression analysis of bovine blastocysts produced by parthenogenic activation or fertilisation

2011 ◽  
Vol 23 (4) ◽  
pp. 591 ◽  
Author(s):  
Rémi Labrecque ◽  
Marc-André Sirard
Keyword(s):  
Gene Expression ◽  
Embryonic Development ◽  
Embryo Development ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Cdna Array ◽  
Developmental Competence ◽  
Time Interval ◽  
Bovine Embryo ◽  
Sperm Entry

The processes underlying the very first moments of embryonic development are still not well characterised in mammals. To better define the kinetics of events taking place following fertilisation, it would be best to have perfect synchronisation of sperm entry. With fertilisation occurring during a time interval of 6 to 12 h in the same group of fertilised oocytes, this causes a major variation in the time of activation of embryonic development. Bovine parthenogenesis could potentially result in better synchronisation and, if so, would offer a better model for studying developmental competence. In the present study, bovine oocytes were either parthenogenetically activated or fertilised and cultured in vitro for 7 days. Gene expression analysis for those two groups of embryos at early and expanded stages was performed with BlueChip, a customised 2000-cDNA array developed in our laboratory and enriched in clones from various stages of bovine embryo development. The microarray data analysis revealed that only a few genes were differentially expressed, showing the relative similarity between those two kinds of embryos. Nevertheless, the fact that we obtained a similar diversity of developmental stages with parthenotes suggests that synchronisation is more oocyte-specific than sperm entry-time related. We then analysed our data with Ingenuity pathway analysis. Networks of genes involved in blastocyst implantation but also previous stages of embryo development, like maternal-to-embryonic transition, were identified. This new information allows us to better understand the regulatory mechanisms of embryonic development associated with embryo status.

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