216 COMPARISON BETWEEN A TRI-GAS THERMO INCUBATOR AND A MODULAR CHAMBER WITH PREMIXED GAS ON THE DEVELOPMENT OF BOVINE EMBRYOS IN VITRO

2009 ◽  
Vol 21 (1) ◽  
pp. 206
Author(s):  
M. B. Raito ◽  
M. L. Mphaphathi ◽  
L. M. Schwalbach ◽  
J. P. C. Greyling ◽  
T. L. Nedambale
Keyword(s):  
South African ◽  
Statistical Difference ◽  
Domestic Animals ◽  
Blastocyst Stage ◽  
Bovine Embryos ◽  
Cleavage Rate ◽  
Department Of Agriculture ◽  
Development In Vitro ◽  
Oviductal Fluid

In an attempt to optimize germplasm and reproduction biotechnology IVF laboratory conditions in South Africa, we compared the effects of 2 triple-gas incubation systems, a tri-gas thermo incubator and a modular chamber with premixed gas, on the development of bovine embryos in vitro. After aspirating ovaries collected from a local abattoir, 778 oocytes were matured for 24 h in M-199 supplemented with 10% FBS, and 1 μg mL–1 of FSH and LH at 39°C in 5% CO2. Oocytes were then fertilized in vitro in Brackett and Oliphant (BO) medium at 39°C in 5% CO2. Presumptive zygotes were randomly allocated to the tri-gas thermo incubator or the modular chamber with premixed gas and cultured in synthetic oviductal fluid (SOF) medium at 39°C in 5% CO2, O2, and 90% N2. Total cleavage (Day 2), 8-cell (Day 2), morula (Day 6), and blastocyst (Day 7) rates were recorded postfertilization. Data were analyzed by ANOVA. There was no statistical difference in total cleavage rate between the 2 incubation systems. However, the 8-cell, morula, and blastocyst rates were significantly higher for the modular chamber group compared with the tri-gas incubator group (Table 1). In summary, this study suggests that the modular chamber with premixed gas was a better system for culturing zygotes of South African domestic animals to the blastocyst stage. Table 1.Effect of modular chamber and tri-gas incubator on embryo development in vitro This work was funded by the South African National Department of Agriculture, DST-PDP, and the National Research Foundation (NRF, Grant Nos. RT21 and 24000).

208 EFFECT OF GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR ON BLASTOCYST DEVELOPMENT AND POST-TRANSFER SURVIVAL OF IN VITRO-PRODUCED BOVINE EMBRYOS

2008 ◽  
Vol 20 (1) ◽  
pp. 183 ◽  
Author(s):  
B. Loureiro ◽  
L. Bonilla ◽  
G. Entrican ◽  
P. J. Hansen
Keyword(s):  
Blastocyst Stage ◽  
Bovine Embryos ◽  
Cho Cell ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Cell Supernatant

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that has been implicated in preimplantation embryo development. Granulocyte-macrophage-CSF improves the proportion of bovine embryos that become blastocysts in vitro (Moraes and Hansen 1997 Biol. Reprod. 57, 1060–1065) and increases blastocyst cell numbers in mice (Robertson et al. 2001 Biol. Reprod. 64, 1206–1215). The long-term goal of the present research was to evaluate the effects of GM-CSF on post-transfer survival of bovine embryos. The experiments used recombinant ovine GM-CSF produced in transfected Chinese hamster ovary (CHO) cells or an equivalent volume of cytokine-free CHO cell supernatant (control). The objective of the first study was to evaluate the effects of GM-CSF on post-transfer survival. Embryos were cultured with 10 ng mL–1 of either GM-CSF or cytokine-free CHO cell supernatant added to culture medium at Day 1 after insemination. Embryos were transferred at Day 7 to lactating dairy cows according to a timed embryo transfer protocol. Pregnancy was evaluated at approximately Day 45 of gestation. There was no significant difference in the proportion of embryos becoming blastocysts at Day 7 after insemination (34.8 v. 37.5% for the control and GM-CSF; SEM = 2.4%). There was also no difference in pregnancy rates between cows receiving control embryos (6/24; 25%) and cows receiving embryos treated with GM-CSF (8/35; 23%). A second study determined the effects of various concentrations of GM-CSF on the development of in vitro-produced embryos to the blastocyst stage. Embryos were cultured in 5% (v/v) oxygen (low oxygen) or atmospheric oxygen (21%, w/v; high oxygen) in the presence of 0, 1, 10, or 100 ng mL–1 of GM-CSF or an equivalent volume of cytokine-free CHO cell supernatant (control). The GM-CSF was added on either Day 1 or Day 5 after insemination. Cleavage rate was accessed on Day 3 after insemination. Stage of development was recorded at Day 7 and Day 8 after insemination. There was no effect of GM-CSF on cleavage rate. Addition of GM-CSF at Day 5 to embryos cultured in low or high oxygen increased the percentage of oocytes that became blastocysts at Day 7 (P < 0.01) and Day 8 (P < 0.01), but addition at Day 1 did not have a significant effect on blastocyst development. The greatest effects of GM-CSF occurred at a concentration of 10 ng mL–1. At this concentration, least squares means for the percentage of oocytes that became blastocysts at Day 7 were 13.9 v. 21.6% (control v. GM-CSF) when GM-CSF was added at Day 5, and 19.5 v. 21.5% when GM-CSF was added at Day 1. The percentage of blastocysts at Day 8 was 20.9 v. 28.7% when GM-CSF was added at Day 5, and 26.7 v. 27.5% when GM-CSF was added at Day 1. In conclusion, GM-CSF can affect the competence of embryos to develop to the blastocyst stage, but at the concentrations and times given, there was no evidence that GM-CSF enhanced embryo survival after transfer.

scholarly journals 273EFFECT OF EPIDERMAL GROWTH FACTOR (EGF) DURING OOCYTE MATURATION ON IN VITRO PRODUCTION OF BOVINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 257
Author(s):  
H.J. Hernandez-Fonseca ◽  
R. Palomares-Naveda ◽  
E. Soto-Belloso ◽  
R. Gonzalez-Fernandez ◽  
A.D. De Ondiz-Sanchez ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Formation Rate ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Cleavage Rate ◽  
Medium Components ◽  
Development In Vitro

Medium components during in vitro maturation (IVM) can significantly influence oocyte maturation and subsequent embryo development in vitro (Rose TA and Bavister BD 1992 Mol. Reprod. Dev. 31, 72–77; Harper K and Brackett B 1993 Biol. Reprod. 48, 409–416). The aim of this experiment was to evaluate the effect of EGF during IVM on further development of bovine embryos in vitro. Bovine ovaries were obtained at a slaughterhouse. Cumulus-oocyte complexes (COC) were aspirated from follicles 2–5mm in diameter. COC were incubated for 24h in either of 3 maturation media: T1 (n=72): modified TCM-199; T2 (n=45): modified TCM-199 supplemented with 10ngmL−1 of EGF;; or T3 (n=46): modified TCM-199 supplemented with 10% fetal bovine serum (FBS). After 24h of IVM, COC were inseminated with 2×106 motile spermatozoa/ml. After 18h of gamete coincubation, presumptive zygotes were denuded and placed in culture in SOF rich in glutamine (g-SOf) for 72h, at which time, cleavage rate (%) wass assesed (embryos with &gt;4 cells). Subsequently, cleaved embryos were incubated for an additional 72h in c-SOF (SOF rich in citrate and glucose). Finally, embryos were cultured in modified TCM-199 for 24–48h, at which time blastocyst formation rate (%) was evaluated. Cleavage rates were similar between T2 and T3 but significantly greater than in T1 (P&gt;0.05; see Table 1). Addition of EGF during IVM (T2;; 11/45, 24.4%) did not yield more blastocysts compared to the other two treatments (6/57, 10.5% and 10/29, 34.5%, T1 and T3, respectively). Nonetheless, T3 (with serum) had a greater yield of blastocysts compared to T1 (P&gt;0.01). Results in this study show that the addition of EGF to chemically defined media results in similar cleavage rates and blastocyst yields to those obtained when using serum during IVM. Key words: in vitro maturation, EGF, cleavage, bovine, embryo. Table 1 Effect of EGF and serum during IVM on cleavage rate of bovine oocytes

scholarly journals 245RELATIVE ABUNDANCE OF HSP 70 MRNA IN BOVINE EMBRYOS PRODUCED IN VITRO USING DIFFERENT EMBRYO/VOLUME RATIOS IN CULTURE

2004 ◽  
Vol 16 (2) ◽  
pp. 243
Author(s):  
A.T.D. Oliveira ◽  
C. Gebert ◽  
R.F.F. Lopes ◽  
H. Niemann ◽  
J.L. Rodrigues
Keyword(s):  
Relative Abundance ◽  
Blastocyst Stage ◽  
Culture Conditions ◽  
Hatching Rate ◽  
Bovine Embryos ◽  
Blastocyst Formation ◽  
Cleavage Rate ◽  
Gene Transcripts

In spite of in vitro embryo production systems having been greatly improved over recent years, employing a variety of culture conditions (media, protein sources, gas atmosphere, etc.), we still do not know much about the real necessity of embryos to develop under the same conditions as occur in vivo. These differences between in vivo and in vitro culture at preimplantation embryonic stages can produce deviations in gene expression and in normal fetal development (large offspring syndrome). Heat shock proteins (Hsp) are engaged in cell response to regulatory signals or perturbations in the microenviroment and can be used as a sensitive indicator of stress caused by suboptimal culture conditions (Wrenzycki et al., 2001Hum. Reprod. 16, 893–901). Hsp act as chaperones in facilitating protein folding and assembly and stabilize damaged proteins to prevent aggregation of fragments, thereby allowing repair or degradation. The aim of the present study was to investigate the effects of different embryo/volume ratios on bovine embryo development and the relative abundance of Hsp 70.1 gene transcripts. In this experiment, oocytes were isolated from slaugterhouse ovaries and matured, fertilized and cultured in groups of 5, 10, 20 or 30 per each drop of 100μL. The oocytes were matured in TCM 199 supplemented with 0.4% BSA. After maturation, oocytes were fertilized in TALP medium, using frozen/thawed sperm, selected using a percoll density gradient. The zygotes were cultured to the morula or Day 7 blastocyst stage employing SOF supplemented with 0.4 % BSA. Developmental check points were cleavage rate (Day 3pi), blastocyst formation (Day 8pi) and hatching (Day 11pi). A semi-quantitative RT-PCR assay was used to determine the relative levels of gene transcripts in single embryos at morula (Day 6) and blastocyst (Day 7) stages (Wrenzycki et al., 2001 Biol. Reprod. 65, 309–317). Data of cleavage, blastocyst formation and hatching rates were analyzed using chi-square test. Relative abundance (RA) of Hsp 70.1mRNA were compared in tested groups using ANOVA followed a Tukey test. Differences at P&lt;0.05 were considered significant. Results show that no significative difference in hatching rate per blastocyst produced was detected among the four groups. Cleavage rate and blastocyst formation were significantly higher in groups with 5, 10 and 20 embryos compared with drops containing 30 embryos. Hsp transcripts were detected in morula and blastocyst stages in all groups. In morula stage, no differences were observed in the RA of Hsp 70.1mRNA among groups with 5, 10, 20 and 30 embryos cultured per drop. However, in blastocyst stage, the RA was significantly increased in the group with 20 embryos per drop as compared to the group with 5 embryos. The results show that different embryo/volume ratios in culture influence not only cleavage rate, blastocyst formation and hatching rate, but also expression of Hsp 70.1 gene. Further studies changing other culture conditions and using in vivo-derived bovine embryos will aid in elucidating which culture systems are ideal to produce bovine embryos in vitro. This research was supported by CAPES/DAAD program and CNPq.

214 EFFECT OF OOCYTE SOURCE ON THE DEVELOPMENTAL CAPACITY OF SOUTH AFRICAN IN VITRO-PRODUCED INDIGENOUS CATTLE EMBRYOS

2009 ◽  
Vol 21 (1) ◽  
pp. 205
Author(s):  
T. L. Nedambale ◽  
M. B. Raito ◽  
M. L. Mphaphathi
Keyword(s):  
Treatment Group ◽  
South African ◽  
Formation Rate ◽  
Developmental Rate ◽  
Bovine Embryo ◽  
Department Of Agriculture ◽  
Indigenous Cattle ◽  
Developmental Capacity ◽  
Development In Vitro

The present study was undertaken to determine whether the source of oocytes (slaughterhouse ovaries from feedlot cows or naturally grazing indigenous cows) would affect in vitro bovine embryo production. Bovine oocytes (n = 1047), aspirated from slaughterhouse ovaries from feedlot cows and naturally grazing indigenous cows were randomly allocated to Sanyo, Forma, and Thermo 5% CO2 incubators. Oocytes were then in vitro matured in TCM-199 plus 10% fetal bovine serum, 1 μg mL–1 for both FSH and LH at 39°C for 24 h, and fertilized in Brackett and Oliphant (BO) medium per treatment group at 39°C. Presumptive zygotes were cultured in vitro per treatment group. Total cleavage and blastocyst rates were recorded postfertilization. Data were analyzed by ANOVA. Preliminary results demonstrated that there was no effect of incubator or source of oocytes on cleavage and 8-cell embryos. However, the cleavage and embryo developmental rate tended to be lower for the feedlot group, regardless of the incubator used (Table 1). In conclusion, this study suggests that slaughterhouse ovaries obtained from South African indigenous cows from a feedlot resulted in a lower blastocyst formation rate. Further studies are currently underway to count the cell numbers and to conduct embryo transfer. Table 1.Comparison of three different incubators and source of oocytes on embryo development in vitro This work was funded by the South African National Department of Agriculture, DST-PDP, and the National Research Foundation (NRF, Grant. Nos. RT21 and 24000).

Effect of α-tocopherol on in vitro culture of bovine embryos

2007 ◽  
Vol 87 (4) ◽  
pp. 539-542 ◽  
Author(s):  
A. Marques ◽  
G. Antunes ◽  
P. Santos ◽  
A. Chaveiro ◽  
F. Moreira da Silva
Keyword(s):  
Culture Media ◽  
Blastocyst Stage ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Blastocyst Development ◽  
Media Control ◽  
Oviductal Fluid ◽  
Bovine Oocytes ◽  
Key Words Antioxidant

Bovine oocytes were matured and fertilised under 5% CO2. Presumptive zygotes were co-cultured in synthetic oviductal fluid droplets, supplemented with either 0 (control), 25, 50, 100, or 200 µM of α-tocopherol. Blastocyst development rates were significantly influenced (P < 0.05) by the level of antioxidant in the culture media. Control showed a blastocyst yield of 18.46, 21.11, 27.92, and 31.66%, respectively, at α25; α50 and α100. Blastocyst yield for α200 was severely decreased, to 8.01%. An increase in overall IVF results was also observed as the concentration of α-tocopherol increased (control, 14.21%; α25, 14.35%; α50, 19.52%; α100, 21.11%), decreasing to 5.75% for the concentration of α200. The present study clearly demonstrates that α-tocopherol at a concentration of 100 µM significantly improves the proportion of oocytes that develop to the blastocyst stage. Key words: Antioxidant, α-tocopherol, reactive oxygen species, bovine, embryo

scholarly journals Embryo culture in presence of oviductal fluid induces DNA methylation changes in bovine blastocysts

Reproduction ◽  
2017 ◽  
Vol 154 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Antonio D Barrera ◽  
Elina V García ◽  
Meriem Hamdi ◽  
María J Sánchez-Calabuig ◽  
Ángela P López-Cardona ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Blastocyst Stage ◽  
Early Embryo ◽  
Cpg Methylation ◽  
Bovine Embryos ◽  
Cell Stage ◽  
The One ◽  
Genomic Regions ◽  
Oviductal Fluid

During the transit through the oviduct, the early embryo initiates an extensive DNA methylation reprogramming of its genome. Given that these epigenetic modifications are susceptible to environmental factors, components present in the oviductal milieu could affect the DNA methylation marks of the developing embryo. The aim of this study was to examine if culture of bovine embryos with oviductal fluid (OF) can induce DNA methylation changes at specific genomic regions in the resulting blastocysts. In vitro produced zygotes were cultured in medium with 3 mg/mL bovine serum albumin (BSA) or 1.25% OF added at the one- to 16-cell stage (OF1–16), one- to 8-cell stage (OF1–8) or 8- to 16-cell stage (OF8–16), and then were cultured until Day 8 in medium with 3 mg/mL BSA. Genomic regions in four developmentally important genes (MTERF2, ABCA7, OLFM1, GMDS) and within LINE-1 retrotransposons were selected for methylation analysis by bisulfite sequencing on Day 7–8 blastocysts. Blastocysts derived from OF1–16 group showed lower CpG methylation levels in MTERF2 and ABCA7 compared with the BSA group. However, CpG sites within MTERF2, ABCA7 and OLFM1 showed higher methylation levels in groups OF1–8 and OF8–16 than in OF1–16. For LINE-1 elements, higher CpG methylation levels were observed in blastocysts from the OF1–16 group than in the other experimental groups. In correlation with the methylation changes observed, mRNA expression level of MTERF2 was increased, while LINE-1 showed a decreased expression in blastocysts from OF1–16 group. Our results suggest that embryos show transient sensitivity to OF at early stages, which is reflected by specific methylation changes at the blastocyst stage.

scholarly journals Phyto-oestrogens affect fertilisation and embryo development in vitro in sheep

2018 ◽  
Vol 30 (8) ◽  
pp. 1109 ◽  
Author(s):  
Anna Aryani Amir ◽  
Jennifer M. Kelly ◽  
David O. Kleemann ◽  
Zoey Durmic ◽  
Dominique Blache ◽  
...  
Keyword(s):  
Embryo Development ◽  
Reproductive Function ◽  
Blastocyst Stage ◽  
Biochanin A ◽  
Detectable Effect ◽  
Cleavage Rate ◽  
Cell Counts ◽  
Blastocyst Rate ◽  
Development In Vitro

Phyto-oestrogens such as isoflavones are natural compounds that can profoundly affect reproductive function. In the present study, we tested whether including isoflavone compounds (genistein, biochanin A, formononetin) in the maturation medium would affect the outcomes for ovine oocytes in vitro. Each isoflavone compound was evaluated at five concentrations (0, 2.5, 5, 10, 25 µg mL−1) and the entire protocol was repeated four times. Cumulus–oocyte complexes were randomly allocated to the treatments, then fertilised and cultured in vitro. Compared with control (0 µg mL−1), the lower concentrations of isoflavone (2.5, 5 and 10 µg mL−1) had no detectable effect on the rates of cleavage or embryo development, or on embryo total cell counts (TCC). However, the highest concentration (25 µg mL−1) of all three isoflavones exerted a variety of effects (P < 0.05): genistein decreased cleavage rate, blastocyst rate and blastocyst efficiency (blastocysts produced per 100 oocytes); biochanin A decreased cleavage rate and blastocyst efficiency; and formononetin decreased blastocyst rate and blastocyst efficiency. Biochanin A (25 µg mL−1) reduced embryo TCC specifically at the hatched blastocyst stage (P < 0.05). We conclude that the presence of isoflavones at 25 µg mL−1 during IVM decreases the cleavage rate and inhibits blastocyst hatching.

scholarly journals Development of cultured bovine embryos after exposure to high temperatures in the physiological range

Reproduction ◽  
2001 ◽  
pp. 107-115 ◽  
Author(s):  
RM Rivera ◽  
PJ Hansen
Keyword(s):  
Heat Shock ◽  
Embryonic Development ◽  
High Temperatures ◽  
Deleterious Effect ◽  
Blastocyst Stage ◽  
The Body ◽  
Body Temperatures ◽  
Cleavage Rate ◽  
Development In Vitro

Embryonic development is inhibited by exposure of cultured embryos to high temperatures. However, culture temperatures used to demonstrate the effects of heat on development have been higher than the body temperatures experienced typically by heat-stressed cows. The aim of this study was to determine whether exposing bovine oocytes and embryos to temperatures characteristic of body temperatures of heat-stressed cows would affect embryonic development in vitro. The CO2 percentage of the gas phase was adjusted in all experiments to prevent pH changes in the medium caused by decreased solubility of CO2 at high temperatures. Fertilization of oocytes at 41.0 degrees C reduced cleavage rate and the percentage of oocytes that became blastocysts compared with at 38.5 degrees C. There was no deleterious effect of fertilization at 40.0 degrees C. When putative zygotes and two-cell embryos were exposed to a range of temperatures from 38.5 to 41.0 degrees C for 3, 6, 9 or 12 h, heat shock reduced the number that developed to the blastocyst stage but only after exposure to 41.0 degrees C for 9 or 12 h. In addition, it was tested whether low O2 tension would reduce the detrimental effects of heat shock. The deleterious effect of 41.0 degrees C was not dependent upon oxygen content or the gas mixture used for culture (5% versus 20.95% O2), indicating that the deleterious effects of heat shock did not depend upon a high O2 environment. In the final experiment, embryos were exposed to 24 h fluctuations in temperature designed to mimic the rectal temperatures of cows exposed to heat stress. Exposure of embryos to this pattern of temperatures starting after fertilization reduced development when embryos were exposed to this environment for 8 days but not when embryos were exposed for 1 day only. These findings indicate that embryonic development can be disrupted by a short-term severe or a prolonged mild heat shock and that the effects of heat shock are not artefacts of changes in pH or high oxygen tension.

219 IN VITRO FERTILIZATION RATE OF MATURED PIG OOCYTES BY FROZEN - THAWED KOLBROEK PIG SPERM CELLS

2011 ◽  
Vol 23 (1) ◽  
pp. 208
Author(s):  
M. H. Mapeka ◽  
K. C. Lehloenya ◽  
M. L. Mphaphathi ◽  
T. L. Nedambale
Keyword(s):  
In Vitro Fertilization ◽  
South African ◽  
Fetal Bovine Serum ◽  
Fertilization Rate ◽  
Blastocyst Stage ◽  
Sperm Cells ◽  
Department Of Agriculture ◽  
Vitro Fertilization ◽  
Fetal Bovine

No studies have investigated the IVF rate of South African indigenous Kolbroek sperm cells following cryopreservation. The objective of this study was to test if frozen–thawed Kolbroek pig sperm cells could penetrate pig oocytes matured in vitro. Pig ovaries were collected from a local abattoir and cumulus–oocytes complexes were obtained by aspiration and were then in vitro matured in TCM-199 supplemented with 10% pig follicular fluid, 10% fetal bovine serum, and 1 μg mL–1 of FSH and LH. Following 44 h of incubation, 200 matured pig oocytes were randomly assigned to 2 treatments with frozen–thawed and fresh (control) Kolbroek pig sperm cells. For IVF, Kolbroek sperm cells were in vitro capacitated using Brackett and Oliphant’s sperm wash medium. Matured pig oocytes and sperm cells were co-incubated for 24 h in Brackett and Oliphant’s IVF medium. Following fertilization, presumptive zygotes were in vitro cultured at 39°C in 5% CO2, 5% O2, and 90% N2. Rate of fertilization was identified by the number of cleaved zygotes. Data were analysed by ANOVA. The total motility of Kolboek pig sperm cells used for IVF was 40% for frozen–thawed sperm cells and 80% for fresh sperm cells. The results showed that Kolbroek pig sperm cells were able to penetrate pig oocytes in vitro. However, no significant (P < 0.05) difference was observed in the percentage of cleavage of pig oocytes fertilized with either frozen–thawed (13.25%) or fresh (13.0%) Kolbroek pig sperm cells. The percentage of embryos that developed to the morulae stage was 2% in frozen–thawed sperm cells and was 0% in fresh Kolbroek sperm cells. Furthermore, oocytes fertilized with Kolboek sperm cells did not develop to the blastocyst stage in either treatment. In conclusion, this study demonstrated that frozen–thawed Kolbroek sperm cells are able to fertilize matured pig oocytes in vitro. This study was funded by the Department of Agriculture Forestry and Fishery, ARC, DST-PDP (RT19000), and National Research Foundation (NRF, Grant No. RT21 and 24000).

scholarly journals Nobiletin enhances the development and quality of bovine embryos in vitro during two key periods of embryonic genome activation

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Karina Cañón-Beltrán ◽  
Yulia N. Cajas ◽  
Serafín Peréz-Cerezales ◽  
Claudia L. V. Leal ◽  
Ekaitz Agirregoitia ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Signaling Pathway ◽  
Akt Signaling ◽  
Mitochondrial Activity ◽  
Bovine Embryos ◽  
Akt Signaling Pathway ◽  
Cell Stage ◽  
Development In Vitro

AbstractIn vitro culture can alter the development and quality of bovine embryos. Therefore, we aimed to evaluate whether nobiletin supplementation during EGA improves embryonic development and blastocyst quality and if it affects PI3K/AKT signaling pathway. In vitro zygotes were cultured in SOF + 5% FCS (Control) or supplemented with 5, 10 or 25 µM nobiletin (Nob5, Nob10, Nob25) or with 0.03% dimethyl-sulfoxide (CDMSO) during minor (2 to 8-cell stage; MNEGA) or major (8 to 16-cell stage; MJEGA) EGA phase. Blastocyst yield on Day 8 was higher in Nob5 (42.7 ± 1.0%) and Nob10 (44.4 ± 1.3%) for MNEGA phase and in Nob10 (61.0 ± 0.8%) for MJEGA phase compared to other groups. Mitochondrial activity was higher and lipid content was reduced in blastocysts produced with nobiletin, irrespective of EGA phase. The mRNA abundance of CDK2, H3-3B, H3-3A, GPX1, NFE2L2 and PPARα transcripts was increased in 8-cells, 16-cells and blastocysts from nobiletin groups. Immunofluorescence analysis revealed immunoreactive proteins for p-AKT forms (Thr308 and Ser473) in bovine blastocysts produced with nobiletin. In conclusion, nobiletin supplementation during EGA has a positive effect on preimplantation bovine embryonic development in vitro and corroborates on the quality improvement of the produced blastocysts which could be modulated by the activation of AKT signaling pathway.

Sign in / Sign up

Export Citation Format

Share Document