De novo transcription of thyroid hormone receptors is essential for early bovine embryo development in vitro

2018 ◽  
Vol 30 (5) ◽  
pp. 779 ◽  
Author(s):  
N.-Y. Rho ◽  
F. A. Ashkar ◽  
T. Revay ◽  
P. Madan ◽  
G.-J. Rho ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Embryo Development ◽  
De Novo ◽  
Preimplantation Embryos ◽  
Embryo Morphology ◽  
Bovine Embryo ◽  
Cell Stage ◽  
Protein Levels ◽  
Development In Vitro

Thyroid hormone receptor (THR) α and THRβ mediate the genomic action of thyroid hormones (THs) that affect bovine embryo development. However, little is known about THRs in the preimplantation embryo. The aim of the present study was to investigate the importance of THRs in in vitro preimplantation bovine embryos. THR transcripts and protein levels were detected in developing preimplantation embryos up to the blastocyst stage. Embryonic transcription of THRs was inhibited by α-amanitin supplementation, and both maternal and embryonic transcription were knocked down by short interference (si) RNA microinjection. In the control group, mRNA and protein levels of THRs increased after fertilisation. In contrast, in both the transcription inhibition and knockdown groups there were significant (P < 0.05) decreases in mRNA expression of THRs from the 2-cell stage onwards. However, protein levels of THRs were not altered at 2-cell stage, although they did exhibit a significant (P < 0.05) decrease from the 4-cell stage. Moreover, inhibition of de novo transcripts of THRs using siRNA led to a significant (P < 0.01) decrease in the developmental rate and cell number, as well as inducing a change in embryo morphology. In conclusion, THRs are transcribed soon after fertilisation, before major activation of the embryonic genome, and they are essential for bovine embryo development in vitro.

Nobiletin-induced partial abrogation of deleterious effects of AKT inhibition on preimplantation bovine embryo development in vitro

2021 ◽  
Author(s):  
Yulia N Cajas ◽  
Karina Cañón-Beltrán ◽  
Carolina Núñez-Puente ◽  
Alfonso Gutierrez-Adán ◽  
Encina M González ◽  
...  
Keyword(s):  
Embryo Development ◽  
Culture Media ◽  
Cell Number ◽  
Early Embryo ◽  
Bovine Embryo ◽  
Cell Stage ◽  
Developmental Effect ◽  
Promote Cell Survival ◽  
Development In Vitro

Abstract During preimplantational embryo development, PI3K/AKT regulates cell proliferation and differentiation and nobiletin modulates this pathway to promote cell survival. Therefore, we aimed to establish whether, when the AKT cascade is inhibited using inhibitors III and IV, nobiletin supplementation to in vitro culture media during the minor (2 to 8-cell stage, MNEGA) or major (8 to 16-cell stage, MJEGA) phases of EGA is able to modulate the development and quality of bovine embryos. In vitro zygotes were cultured during MNEGA or MJEGA phase in SOF + 5% FCS or supplemented with: 15 μM AKT-InhIII; 10 μM AKT-InhIV; 10 μM nobiletin; nobiletin+AKT-InhIII; nobiletin+AKT-InhIV; 0.03% DMSO. Embryo development was lower in treatments with AKT inhibitors, while combination of nobiletin with AKT inhibitors was able to recover their adverse developmental effect and also increase blastocyst cell number. The mRNA abundance of GPX1, NFE2L2, and POU5F1 was partially increased in 8- and 16-cell embryos from nobiletin with AKT inhibitors. Besides, nobiletin increased the p-rpS6 level whether or not AKT inhibitors were present. In conclusion, nobiletin promotes bovine embryo development and quality and partially recovers the adverse developmental effect of AKT inhibitors which infers that nobiletin probably uses another signalling cascade that PI3K/AKT during early embryo development in bovine.

scholarly journals Embryonic disc formation following post-hatching bovine embryo development in vitro

Reproduction ◽  
2020 ◽  
Vol 160 (4) ◽  
pp. 579-589 ◽  
Author(s):  
Priscila Ramos-Ibeas ◽  
Ismael Lamas-Toranzo ◽  
Álvaro Martínez-Moro ◽  
Celia de Frutos ◽  
Alejandra C Quiroga ◽  
...  
Keyword(s):  
Embryo Development ◽  
Blastocyst Stage ◽  
Embryo Morphology ◽  
Bovine Embryo ◽  
Pluripotency Marker ◽  
Embryonic Disc ◽  
Development In Vitro ◽  
Apoptosis And Necrosis ◽  
Disc Formation

Failures during conceptus elongation are a major cause of pregnancy losses in ungulates, exerting a relevant economic impact on farming. The developmental events occurring during this period are poorly understood, mainly because this process cannot be recapitulated in vitro. Previous studies have established an in vitro post-hatching development (PHD) system that supports bovine embryo development beyond the blastocyst stage, based on agarose gel tunnels and serum- and glucose-enriched medium. Unfortunately, under this system embryonic disc formation is not achieved and embryos show notorious signs of apoptosis and necrosis. The objective of this study has been to develop an in vitro system able to support embryonic disc formation. We first compared post-hatching development inside agarose tunnels or free-floating over an agarose-coated dish in serum- and glucose-enriched medium (PHD medium). Culture inside agarose tunnels shaped embryo morphology by physical constriction, but it restricted embryo growth and did not provide any significant advantage in terms of development of hypoblast and epiblast lineages. In contrast to PHD medium, a chemically defined and enriched medium (N2B27) supported complete hypoblast migration and epiblast survival in vitro, even in the absence of agarose coating. Cells expressing the pluripotency marker SOX2 were observed in ~56% of the embryos and ~25% developed embryonic disc-like structures formed by SOX2+ cells. In summary, here we provide a culture system that supports trophectoderm proliferation, hypoblast migration and epiblast survival after the blastocyst stage.

scholarly journals Thyroid hormone supplementation improves bovine embryo development in vitro

2009 ◽  
Vol 25 (2) ◽  
pp. 334-344 ◽  
Author(s):  
F. A. Ashkar ◽  
E. Semple ◽  
C. H. Schmidt ◽  
E. St. John ◽  
P. M. Bartlewski ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Embryo Development ◽  
Bovine Embryo ◽  
Development In Vitro

Effects of amino acids on development in vitro of cleavage-stage bovine embryos into blastocysts

1996 ◽  
Vol 8 (5) ◽  
pp. 835 ◽  
Author(s):  
T Pinyopummintr ◽  
BD Bavister
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Embryo Development ◽  
Culture Medium ◽  
Essential Amino Acids ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Development In Vitro ◽  
Defined Culture

Effects of amino acids on early bovine embryo development in vitro were examined using a chemically-defined, protein-free culture medium. Bovine embryos produced in vitro were cultured from 18 h to 72 h post insemination in a simple medium containing lactate as the only energy source except for the amino acid treatments. Subsequently, embryos were transferred to TCM-199 supplemented with serum for blastocyst development to substantiate their developmental competence. Treatments were: (1) non-essential amino acids from TCM-199 (NEA); (2) essential amino acids from TCM-199 (EA); (3) NEA+EA; (4) Eagle's minimum essential medium amino acids (MEM AA); (5) 11 amino acids present in HECM-6 (11 AA); and (6) 0.2 mM glutamine (GLN). A higher proportion of embryos (percentage of inseminated ova) cleaved to the > or = 8-cell stage by 72 h post insemination in NEA (56.7%), EA (41.2%), 11 AA (40.3%) and GLN (51.1%) than in either NEA+EA (30.0%) or MEM AA (33.1%). However, after transfer to complex medium, embryos that had developed in EA, as well as those in MEM AA or NEA+EA, produced significantly fewer blastocysts (37.1%, 34.4% and 25.6% respectively) than those in NEA (56.7%), GLN (48.9%) or 11 AA (37.7%). The ability of blastocysts to hatch from their zonae pellucidae was also affected by amino acid treatment during cleavage stages. The present study indicated that the addition of NEA or GLN or 11 AA to a chemically-defined culture medium during the cleavage phase of bovine embryo development increases their subsequent ability to reach the blastocyst stage. These data have implications for understanding the nutritional needs of bovine embryos produced in vitro and for optimizing the composition of culture media to support their development.

21 EFFECT OF SEVERAL PARAMETERS ON PARTHENOGENETIC BOVINE EMBRYO DEVELOPMENT IN VITRO

2006 ◽  
Vol 18 (2) ◽  
pp. 119
Author(s):  
S. Arat ◽  
H. Bagis ◽  
A. Tas ◽  
T. Akkoc
Keyword(s):  
Embryo Development ◽  
Development Rate ◽  
Electrical Pulse ◽  
Bovine Embryo ◽  
Cleavage Rate ◽  
The Third ◽  
Parthenogenetic Embryo ◽  
Development In Vitro ◽  
Parthenogenetic Embryos

The activation of oocytes is one of the most important steps for a successful cloning and has great importance on embryo development in vitro. The objective of this study was to examine the different parameters affecting parthenogenetic embryo development in vitro. In the first experiment, two activation protocols were compared to examine the effect of electrical pulse on activation. Bovine oocytes isolated from slaughterhouse ovaries were matured in TCM-199 supplemented with fetal bovine serum (FBS), sodium pyruvate, penicillin/streptomycin, rat insulin-like growth factor (rIGF-1), bovine follicle-stimulating hormone (bFSH), and bovine luteinizing hormone (bLH). A group of oocytes was exposed to a DC pulse of 133 V/500 �m for 25 �s, and then activated by calcium ionophore (5 �M) for 10 min, cytochalasin D (CD) (2.5 �g/mL) + cycloheximide (CHX, 10 �g/mL) for 1 h, and CHX alone for 5 h (Group 1). Another group of oocytes was activated only by chemicals without electrical pulse. Activated oocytes were cultured for 72 h in G1-3 and then 4-6 days in G2-3 medium. In the second experiment, oocytes activated by electrical pulse and chemicals were cultured in Barc medium for 7-9 days or 72 h in G1-3 and then 4-6 days in G2-3 medium. In the third experiment, oocytes activated by electrical pulse and chemicals were cultured for 48 h or 72 h in G1-3 and then 5-7 days or 4-6 days in G2-3 medium. The differences among groups were analyzed by one-way ANOVA after arcsin square transformation. In the first experiment, cleavage rate (75.6%), development rate (37.3%), and blastocyst cell number (78.4 � 3.2) of oocytes activated by electrical pulse was higher than for the group without electrical pulse (28.7%, 8.0%, 59.5 � 4.3, respectively; P < 0.05). This result showed that activation was started more effectively by electrical pulse than by chemicals. In the second experiment, there was no significant difference on cleavage rate between the two groups (66.6%, 65.0%, respectively), and the blastocyst development rate of parthenogenetic embryos cultured in G1-3/G2-3 (36.6%) was higher than in the Barc medium group (16.6%; P < 0.05). This result showed that G1-3/G2-3 medium was more effective for parthenogenetic embryo development than Barc medium. In the third experiment, although significant differences could not be found between the two groups in the development rate of parthenogenetic embryos cultured for a total of 7-9 days (30.8%, 39.2%, respectively), the development rate of embryos cultured for 72 h in G1-3 was higher (26.4%) than for the 48-h group (15%; P < 0.05) on Day 7. This result showed that embryos developed more slowly when cultured for a shorter time in G1-3 medium before transfer to G2-3 medium. This study was supported by a grant from TUBITAK, Turkey (VHAG-1022).

87 COAGULANSIN-A VIA HEAT SHOCK PROTEIN 70 INDUCTION SHOWS BENEFICIAL EFFECTS ON THE DEVELOPMENT OF BOVINE EMBRYOS IN VITRO

2016 ◽  
Vol 28 (2) ◽  
pp. 173
Author(s):  
I. Khan ◽  
K.-L. Lee ◽  
A.-N. Ha ◽  
P.-R. Park ◽  
S.-H. Song ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Embryo Development ◽  
Heat Shock Protein 70 ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Bovine Embryo ◽  
Hsp 70 ◽  
Development In Vitro ◽  
Bovine Oocytes

Coagulansin-A (withanolide) is the steroidal lactone obtained from Withania coagulans, which belong to Solanaceae family. The coagulansin-A induces heat shock protein 70 (HSP-70), which acts as a cellular antioxidant. This study was conducted to investigate the effect of coagulansin-A on bovine oocytes maturation and embryo development in vitro. All these oocytes were aspirated from the ovaries obtained from Korean Hanwoo cows at a local abattoir. To analyse the possible beneficial effect of coagulansin-A on bovine oocytes maturation in vitro, 355 oocytes per group (control and treatment) in seven replicates were subjected with three concentrations i.e. (1, 5, and 10 µM) of coagulansin-A. The coagulansin-A was added in the in vitro-matured (IVM) media for 20 to 22 h followed by IVF for 18 to 22 h, and after fertilization the fertilized oocytes were transferred to IVC1 media for 3 days. After 3 days, the cleavage rate was checked and the 8-cell stage embryos were transferred to IVC2 media and embryo development was checked at Day 8. The culture was carried out at 5% CO2 and 38.5°C. The results indicated that among the three concentrations of Coagulansin-A, only 5 µM remarkably (P < 0.05) improved embryo development (Day 8 blastocyst), being 27.30% and 40.01% for control and treated groups, respectively. This concentration also significantly (P < 0.05) encouraged the activation of HSP-70, having 16.44 arbitrary units (AU) and 35.41 AU integral optical density (IOD) for control and treated groups, respectively. The immunofluorescence analysis revealed that 5 µM coagulansin-A supplementation significantly (P < 0.05) inhibited oxidative stress and inflammation during bovine embryo development in vitro by decreasing IOD of 8-Oxoguanosine (8-OxoG) from 28.12 AU in control to 18.06 AU for the treated group and nuclear factor kappa B (NF-kB) IOD (P < 0.05) from 42.25 AU to 21.80 for control and treated groups, respectively. Additionally, the results obtained from terminal deoxynucleotidyl transferase TUNEL assay confirmed that coagulansin-A treatment reduced the bovine embryo DNA damage significantly (P < 0.05) from 7.4 ± 0.375 to 5.7 ± 0.287 and improved the embryo quality (P < 0.05) with mean cell numbers of 127.7 ± 4.161 and 150.1 ± 3.624 per embryo for control and coagulansin-A treated groups, respectively. This study provides new information regarding the mechanisms by which coagulansin-A promotes bovine oocyte maturation and embryo development in vitro.

scholarly journals Exogenous paraoxonase-1 during oocyte maturation improves bovine embryo development in vitro

2016 ◽  
Vol 51 (5) ◽  
pp. 827-830 ◽  
Author(s):  
JAA Rincón ◽  
EM Madeira ◽  
FT Campos ◽  
B Mion ◽  
JF Silva ◽  
...  
Keyword(s):  
Embryo Development ◽  
Oocyte Maturation ◽  
Paraoxonase 1 ◽  
Bovine Embryo ◽  
Development In Vitro

Effect of oviduct-conditioned medium and of cumulus cells on bovine embryo development in vitro

1992 ◽  
Vol 37 (1) ◽  
pp. 256 ◽  
Author(s):  
P. Mermillod ◽  
C. Boccart ◽  
C. Wils ◽  
A. Massip ◽  
F. Dessy
Keyword(s):  
Conditioned Medium ◽  
Embryo Development ◽  
Cumulus Cells ◽  
Bovine Embryo ◽  
Development In Vitro
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