Saint Hyacinthe, United States
Saint Hyacinthe, United States

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Colleau J.J.,French National Institute for Agricultural Research | Sargolzaei M.,LAlliance Boviteq | Sargolzaei M.,University of Guelph
Journal of Animal Breeding and Genetics | Year: 2011

In real data, inbreeding is usually underestimated because of missing pedigree information. A method adapted to the dairy cattle situation is presented to approximate inbreeding when the stored population pedigree is incomplete. Missing parents in incomplete pedigrees were given a dummy identification and assigned to groups (up to nine for a given birth date of progeny). These groups were linked to contemporary reference groups with known parents. An explicit model considered that polygenic breeding values in a censored group were centred on a function of the average breeding value in the corresponding reference group and deviated independently. Inbreeding coefficients were obtained progressively over birth dates starting from founders. For each date considered, the parameters pertaining to its groups were computed using the parameters already obtained from groups belonging to the previous dates. The updating algorithms were given in detail. An indirect method was implemented to expedite mass computations of the relationship coefficients involved (MIM). MIM was compared to Van Raden's (VR) method using simulated populations with 20 overlapping generations and different rates of missing sires and dams. In the situation of random matings, the average inbreeding coefficients by date obtained by MIM were close to true values, whereas they were strongly underestimated by VR. In the situation of assortative matings, MIM gave average inbreeding coefficients moderately underestimated, whereas those of VR's method were still strongly underestimated. The main conclusion of this study adapted to the situation of dairy cattle with incomplete pedigrees was that corrections for inbreeding and coancestry coefficients are more efficient with an explicit appropriate genetic model than without. © 2011 Blackwell Verlag GmbH.

Gilbert I.,Laval University | Robert C.,Laval University | Dieleman S.,University Utrecht | Blondin P.,LAlliance Boviteq | Sirard M.-A.,Laval University
Reproduction | Year: 2011

The LH surge induces a multitude of events that are essential for ovulation and corpus luteum formation. The transcriptional responses to the LH surge of preovulatory granulosa cells (GCs) are complex and still poorly understood. In this study, a genome-wide bovine oligo array was used to determine how the gene expression profile of GCs is modulated by the LH surge. GCs from three different stages were used to assess the short- and long-term effects of this hormone on follicle differentiation: 1) 2 h before induction of the LH surge, 2) 6 h and 3) 22 h after the LH surge. The results obtained were a list of differentially expressed transcripts for each GC group. To provide a comprehensive understanding of the processes at play, biological annotations were used to reveal the different functions of transcripts, confirming that the LH surge acts in a temporal manner. The pre-LH group is involved in typical tasks such as cell division, development, and proliferation, while the early response to the LH surge included features such as response to stimulus, vascularization, and lipid synthesis, which are indicative of cells preparing for ovulation. The late response of GCs revealed terms associated with protein localization and intracellular transport, corresponding to the future secretion task that will be required for the transformation of GCs into corpus luteum. Overall, results described in this study provide new insights into the different transcriptional steps that GCs go through during ovulation and before luteinization. © 2011 Society for Reproduction and Fertility.

Labrecque R.,Laval University | Vigneault C.,LAlliance Boviteq | Blondin P.,LAlliance Boviteq | Sirard M.-A.,Laval University
Theriogenology | Year: 2014

Ovarian stimulation with FSH combined with an appropriate period of FSH withdrawal (coasting) before ovum pick-up now appears to be a successful way to obtain oocytes with high developmental competence in bovine. Recent results showed that extending follicular growth by only 24hours has a detrimental effect on oocyte quality as shown by the reduced blastocyst formation rate. Although these treatments are initiated during the luteal phase with low LH level, the small LH pulsatility present at that time could potentially impact follicular development as well as oocyte quality. In this study, a GnRH antagonist (Cetrotide) was used to suppress LH secretion during follicular differentiation to get a better insight into the physiological importance of the LH support during that period. Oocytes were collected by ovum pick-up, and quality was assessed by measuring the blastocyst formation rate obtained after IVM-IVF. The oocyte transcriptome from GnRH antagonist-treated animals was also compared with that from a control group (coasting duration of 68hours) to detect possible alterations at the messenger RNA (mRNA) level. The oocyte quality was not statistically affected by the treatment as shown by the blastocyst formation rate obtained. However, microarray analysis showed that a total of 226 genes had a significant difference (fold change > 2; P < 0.05) at the mRNA level, with the majority being in overabundance in the treated group. Many genes related to RNA posttranscriptional modifications presented different abundance at the mRNA level significant differences in the control group (68hours), whereas translation function appeared to be affected, with many genes related to structural constituents of the ribosome presenting a overabundance in the GnRH antagonist-treated group. Specific mRNAs with crucial roles in chromosome segregation control also showed significant difference at the mRNA level after Cetrotide treatment. The results presented here indicated that the suppression of the LH secretion in an optimal stimulated context would have an impact on the oocyte, with the possible alteration of critical functions related to translation capacity and chromosome segregation control. © 2014 Elsevier Inc.

Labrecque R.,Laval University | Vigneault C.,LAlliance Boviteq | Blondin P.,LAlliance Boviteq | Sirard M.-A.,Laval University
Molecular Reproduction and Development | Year: 2013

Recent progress in the ovarian stimulation protocol used for bovine in vitro maturation and fertilization, especially through optimization of the follicle-stimulating hormone (FSH) withdrawal period ("coasting") after ovarian pre-treatment with FSH, has significantly improved blastocyst outcome. Despite this important success, the underlying factors leading to improved oocyte quality have not yet been identified. The aim of this project was to compare the transcriptome of germinal vesicle-stage oocytes collected from FSH-stimulated cows after various coasting periods (20, 44, 68, and 92hr) to determine which transcripts were accumulated or depleted during the rise and fall of competence. Oocytes from each coasting period were compared to the three other times (optimal conditions, 44 and 68hr; under-matured, 20hr; and over-matured, 92hr) per animal, allowing each cow to be its own control (24 collections). Microarray analysis revealed that between 5 and 338 transcripts were significantly different across the six comparisons, with an important longitudinal modulation in terms of gene expression profile. Not surprisingly, as the transcriptional activity decreased in these oocytes, several transcripts that are significantly modulated during coasting are related to RNA processing functions, as shown by functional analysis. Ingenuity Pathway Analysis also highlighted another important function: the control of chromosome segregation. The results presented here indicate that the quality gained with the optimal coasting time does not last, and also suggests a possible mechanism of control by transcript degradation that could be implicated if the oocyte is not ovulated at the right time. © 2013 Wiley Periodicals, Inc.

Gilbert I.,Laval University | Robert C.,Laval University | Vigneault C.,LAlliance Boviteq | Blondin P.,LAlliance Boviteq | Sirard M.-A.,Laval University
Reproduction | Year: 2012

In the case of in vitro embryonic production, it is known that not all oocytes detain the developmental capacity to form an embryo. This capacity appears to be acquired through completion of folliculogenesis, during which the oocyte and follicular cells influence their respective destinies. The differentiation status of granulosa cells (GCs) could therefore offer an indicator of oocyte quality. The aim of this study was to compare mRNA transcript abundance in GCs associated with oocytes that subsequently reach or not the blastocyst stage. GCs were collected from cattle following an ovarian stimulation protocol that did or did not include the administration of LH. GCs were classified according to the developmental stage achieved by the associated oocytes. Transcript abundance was measured by microarray. Follicles (n=189) obtained from cows before and after the LH surge were essentially similar and the rates of oocytes reaching the blastocyst stage were not significantly different (52 vs 41%), but blastocyst quality was significantly better in the post-LH-surge group. In GCs from the pre-LH-surge group and associated with developmentally competent oocytes, 18 overexpressed and 22 underexpressed transcripts were found, including novel uncharacterized transcripts, whereas no differentially expressed transcripts were associated with developmentally different oocytes in the post-LH-surge group. The novel transcriptomic response associated with LH appeared to mask the difference. Based on oocyte developmental competence, the period prior to the LH surge appears best suited for studying competence-associated mRNA transcripts in bovine follicle cells. © 2012 Society for Reproduction and Fertility.

Cagnone G.L.M.,Laval University | Dufort I.,Laval University | Vigneault C.,LAlliance Boviteq | Sirard M.-A.,Laval University
Biology of Reproduction | Year: 2012

To understand the compromised survival of embryos derived from assisted reproductive techniques, transcriptome survey of early embryonic development has shown the impact of in vitro culture environment on gene expression in bovine or other living species. However, how the differentially expressed genes translate into developmentally compromised embryos is unresolved. We therefore aimed to characterize transcriptomic markers expressed by bovine blastocysts cultured in conditions that are known to impair embryo development. As increasing glucose concentrations has been shown to be stressful for early cleavage stages of mammalian embryos and to decrease subsequent blastocyst survival, in vitro-matured/fertilized bovine zygotes were cultured in control (0.2 mM) or high-glucose (5 mM) conditions until the 8- to 16-cell stage, and then transferred to control media until they reached the blastocyst stage. The concentration of 5 mM glucose was chosen as a stress treatment because there was a significant effect on blastocyst rate without the treatment's being lethal as with 10 mM. Microarray analysis revealed gene expression differences unrelated to embryo sex or hatching. Overrepresented processes among differentially expressed genes in treated blastocysts were extracellular matrix signalling, calcium signaling, and energy metabolism. On a pathophysiological level, higher glucose treatment impacts pathways associated with diabetes and tumorigenesis through genes controlling the Warburg effect, i.e., emphasis on use of anaerobic glycolysis rather than oxidative phosphorylation. These results allowed us to conclude that disruption of in vitro preattachment development is concomitant with gene expression modifications involved in metabolic control.

Nivet A.-L.,Laval University | Bunel A.,Laval University | Labrecque R.,Laval University | Belanger J.,LAlliance Boviteq | And 3 more authors.
Reproduction | Year: 2012

Combinations of genetic, environmental, and management factors are suspected to explain the loss in fertility observed for over 20 years in dairy cows. In some cases, IVF is used. When compared with in vivo embryo production, IVF resulted in low success rates until the FSH coasting process (FSH starvation after superstimulation) was introduced in 2002. Increased competence associated with FSH withdrawal of aspirated oocyte for in vitro maturation and IVF has not been optimized nor explained yet. The goal here was to determine and characterize the optimal oocyte competence acquisition window during the coasting period by determining blastocyst rates and follicular cohort development. Commercial milking cycling cows (n=6) were stimulated with 3 days of FSH (6 X 40 mg NIH Folltropin-V given at 12 h intervals) followed by a coasting period of 20, 44, 68, or 92 h. Each animal was exposed to the four conditions and served as its own control. At the scheduled time, transvaginal aspirations of immature oocytes were performed followed by IVF of half the oocytes. The outcomes were as follows: i) FSH coasting was optimal at a defined period: between 44 and 68 h of coasting; ii) The best estimated coasting duration was ∼54 ± 7 h; iii) Under these conditions, the best statistical blastocyst rate estimation was ∼70%; iv) Between 44 and 68 h of coasting, follicle size group proportions were similar; v) Follicle diameter was not linearly associated with competence. In conclusion, coasting duration is critical to harvest the oocytes at the right moment of follicular differentiation. © 2012 Society for Reproduction and Fertility.

Blondin P.,LAlliance Boviteq
Animal Reproduction | Year: 2014

The current world population is increasing at a fast rate. In order to feed this larger population, food production must increase by 70 percent. Recent reports show a record global production of 58.9 Carcass Equivalent Weight million metric tonnes of beef expected for 2014. It becomes clear that the worldwide agricultural community will have to integrate new technologies to assure the sustainability of global livestock and meat demands. Agriculture has benefited tremendously from the innovation of reproductive technologies such as semen artificial insemination and cryopreservation, embryo transfer and cryopreservation, and in vitro fertilization. Only recently have some developed countries accepted the import and export of frozen IVF embryos and more countries are currently evaluating this. Before 2003, in vitro embryos represented not more than 20% of all embryos produced. After 2003, this jumped to 30 to 39% of all embryos produced, and is increasing. It is clear that South America, and more specifically Brazil, is driving this increase. However, most people in this field would agree that the trend is true for many regions active in this field. International movement of gametes or embryos must be performed in biosecure manners to make certain that pathogenic organisms are controlled and that transmission of infection to recipient animals and progeny is avoided. The embryo transfer industry has adopted appropriate procedures to manage the biosecurity risks and hence mitigate risks of pathogen transmission through international trade of bovine embryos. Techniques for biosecure production of in vivo bovine embryos have been well established. However, as in vitro embryos are relatively new to this business, there are not many papers on the subject of pathogen-interaction with this type of embryo. Certain studies demonstrate that the decontamination of in vitro embryos using recommended procedures is effective for specific pathogens while others have shown that this is not as evident in other conditions. All agree that more research is needed regarding washing protocols for in vitro embryos. It is imperative that the scientific community continues its research to validate current embryo sanitary washing procedures and recommend any modifications that would be necessary for IVF embryos. As embryos are becoming an important component of international trade of bovine genetics, such research must not only continue but augment if key parties want to assure they meet the worldwide rising need of meat and dairy products.

Bohmanova J.,University of Guelph | Sargolzaei M.,LAlliance Boviteq | Schenkel F.S.,University of Guelph
BMC Genomics | Year: 2010

Background: Effectiveness of genomic selection and fine mapping is determined by the level of linkage disequilibrium (LD) across the genome. Knowledge of the range of genome-wide LD, defined as a non-random association of alleles at different loci, can provide an insight into the optimal density and location of single-nucleotide polymorphisms (SNPs) for genome-wide association studies and can be a keystone for interpretation of results from QTL mapping.Results: Linkage disequilibrium was measured by |D'| and r 2between 38,590 SNPs (spaced across 29 bovine autosomes and the X chromosome) using genotypes of 887 Holstein bulls. The average level of |D'| and r 2for markers 40-60 kb apart was 0.72 and 0.20, respectively in Holstein cattle. However, a high degree of heterogeneity of LD was observed across the genome. The sample size and minor allele frequency had an effect on |D'| estimates, however, r 2was not noticeably affected by these two factors. Syntenic LD was shown to be useful for verifying the physical location of SNPs. No differences in the extent of LD and decline of LD with distance were found between the intragenic and intergenic regions.Conclusions: A minimal sample size of 444 and 55 animals is required for an accurate estimation of LD by |D'| and r 2, respectively. The use of only maternally inherited haplotypes is recommended for analyses of LD in populations consisting of large paternal half-sib families. Large heterogeneity in the pattern and the extent of LD in Holstein cattle was observed on both autosomes and the X chromosome. The extent of LD was higher on the X chromosome compared to the autosomes. © 2010 Bohmanova et al; licensee BioMed Central Ltd.

Tscherner A.,University of Guelph | Gilchrist G.,University of Guelph | Smith N.,University of Guelph | Blondin P.,LAlliance Boviteq | And 2 more authors.
Reproductive Biology and Endocrinology | Year: 2014

Background: Oocyte fertilization and successful embryo implantation are key events marking the onset of pregnancy. In sexually reproducing organisms, embryogenesis begins with the fusion of two haploid gametes, each of which has undergone progressive stages of maturation. In the final stages of oocyte maturation, minimal transcriptional activity is present and regulation of gene expression occurs primarily at the post-transcriptional level. MicroRNAs (miRNA) are potent effectors of post-transcriptional gene silencing and recent evidence demonstrates that the miR-34 family of miRNA are involved in both spermatogenesis and early events of embryogenesis. Methods: The profile of miR-34 miRNAs has not been characterized in gametes or embryos of Bos taurus. We therefore used quantitative reverse transcription PCR (qRT-PCR) to examine this family of miRNAs: miR-34a, -34b and -34c as well as their precursors in bovine gametes and in vitro produced embryos. Oocytes were aspirated from antral follicles of bovine ovaries, and sperm cells were isolated from semen samples of 10 bulls with unknown fertility status. Immature and in vitro matured oocytes, as well as cleaved embryos, were collected in pools. Gametes, embryos and ovarian and testis tissues were purified for RNA. Results: All members of the miR-34 family are present in bovine spermatozoa, while only miR-34a and -34c are present in oocytes and cleaved (2-cell) embryos. Mir-34c demonstrates variation among different bulls and is consistently expressed throughout oocyte maturation and in the embryo. The primary transcript of the miR-34b/c bicistron is abundant in the testes and present in ovarian tissue but undetectable in oocytes and in mature spermatozoa. Conclusions: The combination of these findings suggest that miR-34 miRNAs may be required in developing bovine gametes of both sexes, as well as in embryos, and that primary miR-34b/c processing takes place before the completion of gametogenesis. Individual variation in sperm miR-34 family abundance may offer potential as a biomarker of male bovine fertility. © 2014 Tscherner et al.; licensee BioMed Central Ltd.

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