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.

Large-scale transcriptional analysis of bovine embryo biopsies in relation to pregnancy success after transfer to recipients

2006 ◽  
Vol 28 (1) ◽  
pp. 84-96 ◽  
Author(s):  
Ashraf El-Sayed ◽  
Michael Hoelker ◽  
Franca Rings ◽  
Dessie Salilew ◽  
Danyel Jennen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Glucose Metabolism ◽  
Expression Analysis ◽  
Large Scale ◽  
Gene Expression Analysis ◽  
Cdna Array ◽  
Microarray Data Analysis ◽  
Transcriptional Analysis ◽  
Bovine Embryo ◽  
Protein Amino Acid

The purpose of this work is to address the relationship between transcriptional profile of embryos and the pregnancy success based on gene expression analysis of blastocyst biopsies taken prior to transfer to recipients. Biopsies (30–40% of the intact embryo) were taken from in vitro-produced day 7 blastocysts ( n = 118), and 60–70% were transferred to recipients after reexpansion. Based on the success of pregnancy, biopsies were pooled in three groups (each 10 biopsies) namely: those resulting in no pregnancy (G1), resorbed embryos (G2), and those resulting in calf delivery (G3). Gene expression analysis of these groups was performed using home-made bovine preimplantation-specific cDNA array (219 clones) and BlueChip (with ∼2,000 clones). Microarray data analysis results revealed a total of 52 and 58 genes were differentially regulated during comparison between G1 vs. G3 and G2 vs. G3. Biopsies resulted in calf delivery were enriched with genes necessary for implantation (COX2 and CDX2), carbohydrate metabolism (ALOX15), growth factor (BMP15), signal transduction (PLAU), and placenta-specific 8 (PLAC8). Biopsies from embryos resulting in resorption are enriched with transcripts involved protein phosphorylation (KRT8), plasma membrane (OCLN), and glucose metabolism (PGK1 and AKR1B1). Biopsies from embryos resulting in no pregnancy are enriched with transcripts involved inflammatory cytokines (TNF), protein amino acid binding (EEF1A1), transcription factors (MSX1, PTTG1), glucose metabolism (PGK1, AKR1B1), and CD9, which is an inhibitor of implantation. In conclusion, we generated direct candidates of blastocyst-specific genes which may play an important role in determining the fate of the embryo after transfer.

Gene expression analysis of 1,25(OH)2 D3 -dependent differentiation of HL-60 cells: a cDNA array study

2002 ◽  
Vol 118 (4) ◽  
pp. 1065-1070 ◽  
Author(s):  
Hakan Savli ◽  
Yan Aalto ◽  
Bálint Nagy ◽  
Sakari Knuutila ◽  
Seppo Pakkala
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Cdna Array

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

The requirement for protein kinase C delta (PRKCD) during preimplantation bovine embryo development

2016 ◽  
Vol 28 (4) ◽  
pp. 482 ◽  
Author(s):  
Qi-En Yang ◽  
Manabu Ozawa ◽  
Kun Zhang ◽  
Sally E. Johnson ◽  
Alan D. Ealy
Keyword(s):  
Gene Expression ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Embryonic Development ◽  
Embryo Development ◽  
Early Embryonic Development ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Cell Stage ◽  
Kinase C

Protein kinase C (PKC) delta (PRKCD) is a member of the novel PKC subfamily that regulates gene expression in bovine trophoblast cells. Additional functions for PRKCD in early embryonic development in cattle have not been fully explored. The objectives of this study were to describe the expression profile of PRKCD mRNA in bovine embryos and to examine its biological roles during bovine embryo development. Both PRKCD mRNA and protein are present throughout early embryo development and increases in mRNA abundance are evident at morula and blastocyst stages. Phosphorylation patterns are consistent with detection of enzymatically active PRKCD in bovine embryos. Exposure to a pharmacological inhibitor (rottlerin) during early embryonic development prevented development beyond the eight- to 16-cell stage. Treatment at or after the 16-cell stage reduced blastocyst development rates, total blastomere numbers and inner cell mass-to-trophoblast cell ratio. Exposure to the inhibitor also decreased basal interferon tau (IFNT) transcript abundance and abolished fibroblast growth factor-2 induction of IFNT expression. Furthermore, trophoblast adhesion and proliferation was compromised in hatched blastocysts. These observations provide novel insights into PRKCD mRNA expression profiles in bovine embryos and provide evidence for PRKCD-dependent regulation of embryonic development, gene expression and post-hatching events.

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.

scholarly journals Effects of different oocyte retrieval and in vitro maturation systems on bovine embryo development and quality

Zygote ◽  
2014 ◽  
Vol 23 (3) ◽  
pp. 367-377 ◽  
Author(s):  
Sandra Milena Bernal ◽  
Julia Heinzmann ◽  
Doris Herrmann ◽  
Bernd Timmermann ◽  
Ulrich Baulain ◽  
...  
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Developmental Competence ◽  
Bovine Embryo ◽  
Global Methylation ◽  
Oocyte Developmental Competence

SummaryCyclic adenosine monophosphate (cAMP) modulators have been used to avoid spontaneous oocyte maturation and concomitantly improve oocyte developmental competence. The current work evaluated the effects of the addition of cAMP modulators forskolin, 3-isobutyl-1-methylxanthine (IBMX) and cilostamide during in vitro maturation on the quality and yields of blastocysts. The following experimental groups were evaluated: (i) slicing or (ii) aspiration and maturation in tissue culture medium (TCM)199 for 24 h (TCM24slicing and TCM24aspiration, respectively), (iii) aspiration and maturation in the presence of cAMP modulators for 30 h (cAMP30aspiration) and in vivo-produced blastocysts. In vitro-matured oocytes were fertilized and presumptive zygotes were cultured in vitro to assess embryo development. Cleavage, blastocyst formation, blastocyst cell number, mRNA abundance of selected genes and global methylation profiles were evaluated. Blastocyst rate/zygotes for the TCM24aspiration protocol was improved (32.2 ± 2.1%) compared with TCM24slicing and cAMP30aspiration (23.4 ± 1.2% and 23.3 ± 2.0%, respectively, P<0.05). No statistical differences were found for blastocyst cell numbers. The mRNA expression for the EGR1 gene was down-regulated eight-fold in blastocysts that had been produced in vitro compared with their in vivo counterparts. Gene expression profiles for IGF2R, SLC2A8, COX2, DNMT3B and PCK2 did not differ among experimental groups. Bovine testis satellite I and Bos taurus alpha satellite methylation profiles from cAMP30aspiration protocol-derived blastocysts were similar to patterns that were observed in their in vivo equivalents (P > 0.05), while those from the other groups were significantly elevated. It is concluded that retrieval, collection systems and addition of cAMP modulators can affect oocyte developmental competence, which is reflected not only in blastocyst rates but also in global DNA methylation and gene expression patterns.

scholarly journals Development of a cDNA array for chicken gene expression analysis

BMC Genomics ◽  
2005 ◽  
Vol 6 (1) ◽  
Author(s):  
Joan Burnside ◽  
Paul Neiman ◽  
Jianshan Tang ◽  
Ryan Basom ◽  
Richard Talbot ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Cdna Array ◽  
Chicken Gene

scholarly journals Gene expression analysis during pre‐implantation mouse embryo development using cDNA microarray

2007 ◽  
Vol 21 (5) ◽  
Author(s):  
Hyun Jae Kim ◽  
Seung Ku Lee ◽  
Yun‐Yi Cha ◽  
Hyoun Geun Kim ◽  
Hong‐Shik Yun ◽  
...  
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
Expression Analysis ◽  
Cdna Microarray ◽  
Mouse Embryo ◽  
Gene Expression Analysis ◽  
Mouse Embryo Development
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