scholarly journals Temporal expression of factors involved in chromatin remodeling and in gene regulation during early bovine in vitro embryo development

Reproduction ◽  
2007 ◽  
Vol 133 (3) ◽  
pp. 597-608 ◽  
Author(s):  
Serge McGraw ◽  
Christian Vigneault ◽  
Marc-André Sirard
Keyword(s):  
Embryo Development ◽  
Developmental Stages ◽  
Epigenetic Modification ◽  
Chromatin Modification ◽  
Bovine Embryo ◽  
Mrna Levels ◽  
Temporal Expression ◽  
Regulate Gene Expression ◽  
Preimplantation Embryo Development

Distinct epigenetic modification events regulate gene expression and chromatin structure during the period between the immature oocyte and the blastocyst. Throughout this developmental period, important methylation fluctuations occur on genomic DNA and histones. Finding single orcombinations offactors, which are at work during this period is essential to understand the entire epigenetic process. With this in mind, we assessed the precise temporal expression profile, during preimplantation embryo development, of 15 key regulators involved in RNA, DNA or histone methylation, chromatin modification or silencing and transcription regulation. To achieve this, real-time RT-PCR was used to quantify the mRNA levels of ATF7IP, DMAP1, EHMT1, EHMT2, HELLS, JARID1A, JARID1B, JMJD1A, JMJD2A, LSD1, MeCP2, METTL3, PRMT2, PRMT5 and RCOR2, in the oocyte and throughout in vitro bovine embryo development. Our results demonstrate that all the 15 key regulators were present to different degrees in the developmental stages tested, and they can be divided into three different groups depending on their respective mRNA profile.

Quantitative expression analysis of blastocyst-derived gene transcripts in preimplantation developmental stages of in vitro-produced bovine embryos using real-time polymerase chain reaction technology

2004 ◽  
Vol 16 (8) ◽  
pp. 753 ◽  
Author(s):  
Nermin El-Halawany ◽  
Siriluck Ponsuksili ◽  
Klaus Wimmers ◽  
Markus Gilles ◽  
Dawit Tesfaye ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Expression Pattern ◽  
Embryo Development ◽  
Developmental Stages ◽  
Bovine Embryo ◽  
Chain Reaction ◽  
Quantitative Expression ◽  
Polymerase Chain

The main objective of the present study was to analyse the quantitative expression pattern of genes from a subtracted blastocyst transcriptome throughout the preimplantation developmental stages of in vitro-produced bovine oocytes and embryos. For this purpose, Day 5 morula (M) cDNAs were subtracted from Day 7 blastocyst (B) cDNAs (B–M) and used to establish a B–M subtracted cDNA library, as reported previously. From the total generated clones, 19 were analysed quantitatively. The mRNA samples isolated from pools of immature oocytes (n = 150), mature oocytes (n = 150) and two-cell (n = 80), four-cell (n = 40), eight-cell (n = 20), morula (n = 6) and blastocyst (n = 3) embryos were reverse transcribed and subjected to real-time polymerase chain reaction (PCR) using sequence-specific primers and SYBR green as the DNA dye. A relative standard curve method was used to analyse the real-time data taking the morula stage as a calibrator. Applying suppression subtractive hybridisation (SSH), a total of 71 clones, which represent 33 different expressed sequence tags, were generated and available for analysis. Most transcripts were analysed for the first time in bovine embryogenesis. The real-time PCR has validated the results of SSH positively for 84% (16/19) of transcripts, whereas 16% (3/19) showed deviation in the expression pattern from the one seen during SSH. Several transcript-specific expression patterns were observed for genes that play decisive roles in bovine embryogenesis. In addition to identification, accurately quantifying the expression profiles of transcripts during development will pave the way towards understanding the molecular mechanisms of embryogenesis and their potential role in early embryo development. Most importantly, the present study has contributed to the enrichment of bovine embryo gene collection by generating new transcripts involved in bovine embryo development.

Temporally differential protein expression of glycolytic and glycogenic enzymes during in vitro preimplantation bovine embryo development

2018 ◽  
Vol 30 (9) ◽  
pp. 1245 ◽  
Author(s):  
Manuel García-Herreros ◽  
Constantine A. Simintiras ◽  
Patrick Lonergan
Keyword(s):  
Embryo Development ◽  
Glucose Transporter ◽  
Developmental Stages ◽  
Glycogen Synthase ◽  
Differentially Expressed ◽  
Bovine Embryo ◽  
Cell Stage ◽  
Glucose Transporter 1 ◽  
Glut 1

Proteomic analyses are useful for understanding the metabolic pathways governing embryo development. This study investigated the presence of enzymes involved in glycolysis and glycogenesis in in vitro-produced bovine embryos at five developmental stages leading up to blastocyst formation. The enzymes examined were: (1) glycolytic: hexokinase-I (HK-I), phosphofructokinase-1 (PFK-1), pyruvate kinase mutase 1/2 (PKM-1/2), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and (2) glycogenic: glycogen synthase kinase-3 isoforms α/ β (GSK-3α/β). Glucose transporter-1 (GLUT-1) was also analysed. The developmental stages examined were: (1) 2–4-cell, (2) 5–8-cell, (3) 16-cell, (4) morula and (5) expanded blastocyst. The enzymes HK-I, PFK-1, PKM-1/2, GAPDH and GLUT-1 were differentially expressed throughout all stages (P < 0.05). GSK-3α and β were also differentially expressed from the 2–4-cell to the expanded blastocyst stage (P < 0.05) and GLUT-1 was identified throughout. The general trend was that the abundance of PFK1, GAPDH and PKM-1/2 decreased whereas HK-I, phospho-GSK3α (P-GSK3α) and P-GSK3β levels increased as the embryo advanced. In contrast, GLUT-1 expression peaked at the 16-cell stage. These data combined suggest that in vitro bovine embryo metabolism switches from being glycolytic-centric to glycogenic-centric around the 16-cell stage, the developmental window also characterised by embryonic genome activation.

scholarly journals Inverse relationship between autophagy and CTSK is related to bovine embryo quality

Reproduction ◽  
2020 ◽  
Vol 159 (6) ◽  
pp. 757-766
Author(s):  
Ahmed Z Balboula ◽  
Mansour Aboelenain ◽  
Jianye Li ◽  
Hanako Bai ◽  
Manabu Kawahara ◽  
...  
Keyword(s):  
Embryo Development ◽  
Embryo Quality ◽  
Cell Number ◽  
Poor Quality ◽  
Developmental Competence ◽  
Protective Mechanism ◽  
Bovine Embryo ◽  
Preimplantation Embryo Development ◽  
Autophagic Activity

Improving the quality and the developmental competence of in vitro produced (IVP) embryos is an indispensable goal for assisted reproductive technology. Autophagy is a major protective mechanism for intracellular degradation of unnecessary cytoplasmic components. Autophagy ends by the fusion between autophagic vacuoles and lysosomes, allowing the degradation of the cargo by lysosomal enzymes, especially the cathepsins (CTSs). However, it is still unclear how autophagy and cathepsin K (CTSK) relate to embryo development. This study evaluated (1.) the activities of autophagy and CTSK in relation to bovine embryo quality and (2.) the effect of autophagy induction and/or CTSK inhibition on preimplantation embryo development and quality. We show here that good-quality embryos exhibited a greater autophagic activity and less CTSK activity compared to poor-quality embryos. Blastomeres of an individual embryo may vary in their quality. Good quality blastomeres showed an increased autophagic activity and decreased CTSK activity compared to poor-quality blastomeres within the same embryo at different developmental stages. Importantly, induction of autophagy and/or inhibition of CTSK improved the developmental rate (increased blastocyst and hatching rates) and the quality (increased total cell number and decreased the percentage of apoptotic cells) of IVP bovine embryos. These results demonstrate a promising approach to selectively isolate good-quality embryos and improve the efficiency of IVEP of cattle embryos.

Effect of zinc and copper on preimplantation mouse embryo development in vitro and metallothionein levels

Zygote ◽  
1993 ◽  
Vol 1 (3) ◽  
pp. 225-229 ◽  
Author(s):  
Francesca Vidal ◽  
Juan Hidalgo
Keyword(s):  
Embryo Development ◽  
Developmental Stages ◽  
Blastocyst Stage ◽  
Preimplantation Embryos ◽  
Mrna Levels ◽  
Mouse Embryo Development ◽  
Preimplantation Mouse Embryo ◽  
Preimplantation Mouse ◽  
Development In Vitro

The effect of zinc and copper on the in vitro development of mouse preimplantation embryos and on metallothionein (MT) levels was studied by exposing the embryos to 100 μM concentrations of the metals for 24 h at the 1-cell,2-cell, 6-8-cell, morula and blastocyst stages. Zinc affected embryo development in the early but not in the late stages, whereas copper affected it more generally. The combined presence of both metals caused a stronger embryotoxicity. MT levels were measured by radioimmunoassay and were found to be similar at all developmental stages, though possibly higher at the blastocyst stage. The exposure of embryos to zinc and copper increased MT levels significantly only at the blastocyst stage, supporting previously published results on MT mRNA levels.

scholarly journals Short-term exposure to hydrogen peroxide during oocyte maturation improves bovine embryo development

Reproduction ◽  
2010 ◽  
Vol 139 (3) ◽  
pp. 505-511 ◽  
Author(s):  
L Vandaele ◽  
M Thys ◽  
J Bijttebier ◽  
A Van Langendonckt ◽  
I Donnay ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Stress Tolerance ◽  
Embryo Development ◽  
Apoptotic Cell ◽  
Bovine Embryo ◽  
Short Term ◽  
Preimplantation Embryo Development ◽  
Short Term Exposure

Recent studies have shown that short-term exposure of oocytes to a stressor such as hydrostatic pressure or osmotic stress might induce stress tolerance in embryos. The aim of the present study was to investigate the consequences of short-term hydrogen peroxide (H2O2) exposure to bovinein vitromatured cumulus–oocyte complexes (COCs) on subsequent preimplantation embryo development and apoptosis. In the first experiment, mature COCs were incubated in H2O2at concentrations ranging between 0.01 and 100 μmol/l, and subsequently fertilized and cultured. Oocyte incubation with 50–100 μmol/l of H2O2resulted in a significantly higher blastocyst yield (47.3%) in comparison with control medium (31.8%), while apoptotic cell ratio was inversely related with H2O2concentration. In the second experiment, we showed that the stress tolerance after H2O2exposure was not mediated by increased glutathione content in treated oocytes nor by enhanced fertilization or penetration. Further research should concentrate on the potential role of players that have been associated with stress tolerance in somatic cell lines.

scholarly journals 136 EVIDENCE OF A DIRECT EFFECT OF P4 ON IVF-DERIVED BOVINE 8-CELL EMBRYOS

2005 ◽  
Vol 17 (2) ◽  
pp. 219 ◽  
Author(s):  
C.E. Ferguson ◽  
T.R. Davidson ◽  
M.R.B. Mello ◽  
A.S. Lima ◽  
D.J. Kesler ◽  
...  
Keyword(s):  
Embryo Development ◽  
Direct Effect ◽  
Blastocyst Stage ◽  
Control Group ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Direct Role ◽  
Treatment Groups ◽  
And Control

There has been much debate over a direct role for progesterone (P4) in early bovine embryo development. While previous attempts to supplement bovine embryos in vitro with P4 produced results that vary and are often contradictory, this may be a response of administering P4 at inappropriate times. Therefore, the objective of these experiments was to determine if P4 could exert a direct effect on developing IVF-derived bovine embryos when administered at an appropriate time of embryo development. In Exp. I, IVF-derived bovine 8-cell embryos were randomly allotted to treatments: (1) control, CR1aa medium (n = 168); (2) vehicle, CR1aa + ETOH (0.01%) (n = 170); and (3) P4, CR1aa + ETOH + P4 (20 ng/mL in 50-μL droplet) (n = 173). In Exp. II, IVF-derived bovine 8-cell embryos were randomly allotted to treatments: (1) control, CR1aa medium (n = 160); (2) vehicle, CR1aa + DMSO (0.01%) (n = 180); and (3) P4, CR1aa + DMSO (0.01%) + P4 (20 ng/mL in 50-μL droplet) (n = 170). All embryos were evaluated on Days 6 to 9 post-insemination and rates calculated from 8-cell embryos. In Exp. I, ETOH tended to have a detrimental effect with significantly fewer (P < 0.05) embryos (53%) developing to the blastocyst stage on Day 7 compared with the control (62%) and P4 (71%) groups. At Day 7, significantly more embryos cultured in P4 (71%) developed to the blastocyst stage compared with the control group (62%). P4 treatment significantly increased the number of Grade 1 blastocysts (25%) on Day 7 compared with vehicle (15%) and control (17%) groups. At the end of culture, there were also significantly more Day 9 hatched blastocysts in the P4 group (33%) compared with vehicle (22%) and control (21%) groups. Supplementing P4 in the culture medium increased the rate of development, resulting in significantly more blastocysts (8%) on Day 6 and hatched blastocysts (21%) on Day 8 compared with vehicle (3% and 12%) and control (0% and 8%) groups, respectively. In Exp. II, there were no significant differences between treatment groups for Day 7 blastocysts (control 54%, DMSO 61%, P4 57%) and Day 9 hatched blastocysts (control 46%, DMSO 51%, P4 46%). However, there were significantly more Grade 1 blastocysts in the P4 group (22% and 36%) on Days 6 and 8 compared with vehicle (11% and 23%) and control (13% and 23%) groups, respectively. The lack of improvement in Day 7 blastocysts and Day 9 hatched blastocysts rates leads to further uncertainty in understanding the P4 vehicle interactions. In conclusion, the results of these two experiments indicate that P4 can exert a direct effect on the developing IVF-derived bovine embryo; however, due to P4 vehicle interactions; other inert vehicles need to be explored to further evaluate the direct effects of P4 on the developing bovine embryo.

scholarly journals Effects of recombinant OVGP1 protein on in vitro bovine embryo development

2018 ◽  
Vol 64 (5) ◽  
pp. 433-443 ◽  
Author(s):  
Blanca ALGARRA ◽  
Verónica MAILLO ◽  
Manuel AVILÉS ◽  
Alfonso GUTIÉRREZ-ADÁN ◽  
Dimitrios RIZOS ◽  
...  
Keyword(s):  
Embryo Development ◽  
Bovine Embryo

The effect of perfluorohexanesulfonate (PFHxS) on bovine preimplantation embryo development in vitro

2021 ◽  
Vol 350 ◽  
pp. S169-S170
Author(s):  
I. Hallberg ◽  
M. Moberg ◽  
M. Olovsson ◽  
P. Damdimopoulou ◽  
J. Rüegg ◽  
...  
Keyword(s):  
Embryo Development ◽  
Preimplantation Embryo ◽  
Preimplantation Embryo Development ◽  
Development In Vitro

Effects of changing culture medium on preimplantation embryo development in rabbit

Zygote ◽  
2021 ◽  
pp. 1-6
Author(s):  
Haixia Wang ◽  
Wenbin Cao ◽  
Huizhong Hu ◽  
Chenglong Zhou ◽  
Ziyi Wang ◽  
...  
Keyword(s):  
Embryo Development ◽  
Embryo Culture ◽  
Culture Medium ◽  
Cell Number ◽  
Apoptotic Index ◽  
Total Cell ◽  
Toxic Substances ◽  
Total Cell Number ◽  
Preimplantation Embryo Development

Summary Many studies have focused on the optimization of the composition of embryo culture medium; however, there are few studies involving the effect of a culture medium changing procedure on the preimplantation development of embryos. In this study, three groups were designed: a non-renewal group, a renewal group and a half-renewal group. The levels of reactive oxygen species (ROS), apoptotic index, blastocyst ratio and blastocyst total cell number were analyzed in each group. The results showed that the ROS level and the apoptotic index of blastocyst in the non-renewal group were significantly higher than in the renewal group and the half-renewal group (P < 0.05). The blastocyst ratio and blastocyst total cell number were significantly higher in the half-renewal group than that in non-renewal group and the renewal group (P < 0.05). These results demonstrated that the procedure of changing the culture medium influenced ROS level, apoptotic index, blastocyst ratio and total cell number of blastocysts. In addition, the result suggested that changing the culture medium may lead to a loss of important regulatory factors for embryos, while not changing the culture medium may lead to the accumulation of toxic substances. Half-renewal can alleviate the defects of both no renewal and renewal, and benefit embryo development. This study will be of high value as a reference for the optimization of embryo culture in vitro, and is very significant for assisted reproduction.

Sign in / Sign up

Export Citation Format

Share Document