scholarly journals Effect of Ethanol on Parthenogenetic Activation and α-Tocopherol Supplementation during In Vitro Maturation on Developmental Competence of Summer-Collected Bovine Oocytes

2021 ◽  
Vol 43 (3) ◽  
pp. 2253-2265
Author(s):  
Francisco Báez ◽  
Belén Gómez ◽  
Victoria de Brun ◽  
Nélida Rodríguez-Osorio ◽  
Carolina Viñoles
Keyword(s):  
In Vitro Maturation ◽  
Apoptotic Index ◽  
Developmental Competence ◽  
Vitro Fertilization ◽  
Parthenogenetic Activation ◽  
Oocyte Competence ◽  
Maturation Medium ◽  
Bovine Oocytes

The use of α-tocopherol during in vitro maturation (IVM) is an alternative to minimize the adverse effects of heat stress on oocyte competence. However, α-tocopherol is diluted in ethanol, which can induce oocyte parthenogenetic activation (PA). This study aimed to evaluate the role of ethanol concentration on PA and the effect of α-tocopherol supplementation during IVM on the developmental competence and the expression of key genes in blastocysts derived from summer-collected oocytes. All in vitro embryo production was conducted at 5% O2, 5% CO2 at 38.5 °C. Experiment 1: oocytes were cultured with or without 0.05% ethanol. As positive PA control matured oocytes were subjected to 3% or 7% ethanol for 7 min. Oocytes from all groups were placed in fertilization medium (22 h) and culture medium (9 days). Ethanol at 0.05% during IVM did not induce oocyte PA, however, 3% and 7% ethanol were effective parthenogenetic inductors. Experiment 2: oocytes were cultured in maturation medium supplemented with 0, 50, 100 and 200 μM α-tocopherol, diluted in 0.05% ethanol. After in vitro fertilization and embryo culture, we assessed blastocyst apoptotic index and the transcription of a panel of genes. The results showed that supplementation with 100 μM α-tocopherol reduced apoptotic index and increased the expression of SOD2. In conclusion, 100 μM α-tocopherol, diluted in 0.05% ethanol, can be used during IVM to embryonic quality.

scholarly journals LÍQUIDO FOLICULAR EQÜINO NA MATURAÇÃO IN VITRO DE OÓCITOS BOVINOS

2004 ◽  
Vol 9 (1) ◽  
Author(s):  
M.G.L. PINTO ◽  
M.I.B. RUBIN ◽  
C.A.M. SILVA ◽  
T.F. HILGERT ◽  
M.F. SÁ FILHO ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Mineral Oil ◽  
Control Group ◽  
Sodium Pyruvate ◽  
Vitro Fertilization ◽  
Treatment Groups ◽  
Maturation Medium ◽  
Bovine Oocytes

O desenvolvimento embrionário de oócitos bovinos maturados in vitro (MIV) foi avaliado em meio suplementado com líquido folicular eqüino (Lfe). Foram distribuídos 1045 oócitos em 11 repetições formando três grupos tratamentos (T1, T2, T3) e um controle (C). O meio de maturação utilizado foi o TCM-199 acrescido de piruvato de sódio, hormônio folículo estimulante recombinante (rFSHh) e hormônio luteinizante equino (LHe). Suplementou-se esse meio com 10% de soro de égua em estro para o grupo controle e para T1, T2 e T3, o meio foi suplementado com 5, 10, e 20% de LFe, respectivamente. Os oócitos foram maturados in vitro (MIV) por 24h. A fecundação in vitro (FIV) foi realizada em meio Talp-Fert. A MIV e a FIV foram realizadas em estufa a 39ºC com 5% de CO2 em ar e umidade saturada. Os zigotos foram cultivados em meio SOFaaci, sob óleo mineral no interior de bolsas plásticas gaseificadas. As taxas de clivagem e de blastocistos foram observadas diariamente (D), e em D7, foram superiores (P0,05) às do grupo controle. Em D9, a taxa de blastocistos do T2 foi superior (P0,05). O LFe, na concentração de 10% pode ser utilizado, em substituição ao soro de égua em estro para suplementar o meio de MIV de oócitos bovinos. Equine follicular fluid on in vitro maturation of bovine oocytes Abstract Embryo development of bovine oocytes was evaluated using maturation medium supplemented with equine follicular fluid (eFF). One thousand and forty five (1045) oocytes were distributed in 11 replications forming three treatment groups (T1, T2 e T3) and one Control (C). TCM-199 added with sodium pyruvate, rFSHh and LHe was used as maturation medium. This medium was supplemented with 10% estrous mare serum for Control group, and 5, 10, and 20% eFF, respectively, for T1, T2 e T3 groups. In vitro maturation (IVM) of all groups was performed during 24h. In vitro fertilization (IVF) was performed in TALP-FERT medium. IVM and IVF were carried out in an incubator at 39ºC with 5% CO2 in air and saturated humidity. Zygotes were cultured in SOFaaci medium, under mineral oil in gasified bags. Cleavage and blastocyst rates were daily observed (D), and at D7, were higher (P0.05) for those from control group. At D9, blastocyst rate of T2 was higher (P0.05). The eFF, at a 10% concentration, can replace the use of estrous mare serum to supplement the IVM medium of bovine oocytes.

In vitro maturation and glutathione synthesis of porcine oocytes in the presence or absence of cysteamine under different oxygen tensions: role of cumulus cells

2002 ◽  
Vol 14 (3) ◽  
pp. 125 ◽  
Author(s):  
Y. Z. Bing ◽  
Y. Hirao ◽  
K. Iga ◽  
L. M. Che ◽  
N. Takenouchi ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
In Vitro Maturation ◽  
Cumulus Cells ◽  
Vitro Fertilization ◽  
Male Pronucleus ◽  
Nuclear Maturation ◽  
Maturation Medium ◽  
Oxygen Tensions

The present study was conducted to evaluate the effect of cumulus cells on the in vitro maturation (IVM) and glutathione (GSH) synthesis of porcine oocytes cultured in the presence or absence of cysteamine under different oxygen tensions, and on their subsequent male pronucleus formation after in vitro fertilization (IVF). Cumulus-oocyte complexes (COCs) and cumulus-denuded oocytes (DOs) were cultured for 45 h in modified TCM-199 supplemented with or without 150 m cysteamine under a humidified atmosphere of 5% CO2 in air (20%�O2) or 5% CO2, 5% O2 and 90% N2. When cultured in medium supplemented with cysteamine under 20% O2 tension, the rates of COC maturation to the metaphase II (MII) stage were significantly higher than those of DOs (P<0.05). Regardless of the addition of cysteamine and oxygen tension, the rates of male pronucleus formation in COCs after IVM and IVF were significantly higher than in DOs (P<0.05). The GSH content of oocytes was significantly increased by the addition of cysteamine to the maturation medium (P<0.05), with significantly higher GSH content in COCs than in DOs (P<0.05). However, the GSH content of COCs and DOs was not significantly different when cultured in medium without cysteamine. These results indicate that cumulus cells play an important role in nuclear maturation to MII, GSH synthesis in porcine oocytes cultured in the presence of cysteamine, and subsequent male pronucleus formation after IVF.

169 INFLUENCE OF TIME BEFORE BOS INDICUS OOCYTE ASPIRATION ON EMBRYO DEVELOPMENTAL COMPETENCE, EXPRESSION OF MATER AND OCT-4, AND FOLLICULAR STEROID CONCENTRATION

2014 ◽  
Vol 26 (1) ◽  
pp. 198
Author(s):  
R. Urrego ◽  
E. Herrera ◽  
N. Chavarría ◽  
O. Camargo ◽  
N. Rodriguez-Osorio
Keyword(s):  
Follicular Fluid ◽  
Mrna Abundance ◽  
Developmental Competence ◽  
Progesterone Concentration ◽  
Vitro Fertilization ◽  
Oocyte Competence ◽  
Group I ◽  
Group Ii ◽  
Bovine Oocytes

The ability of bovine embryos to develop to the blastocyst stage, to implant, and to generate healthy offspring, depends greatly on the oocyte contribution. Oocyte competence is attributed to its close communication with the follicular environment and to its capacity to synthesise and store great amounts of mRNA. Higher developmental competence of bovine oocytes has been associated with the expression of certain genes and with the steroid concentration in the follicular fluid. Hence, the aim of this study was to establish the influence of OCT-4 and MATER mRNA abundance in the oocyte and the influence of progesterone and oestradiol follicular fluid concentration on the competence of bovine oocytes retrieved 30 min or 4 h after slaughter. Cumulus–oocyte complexes (COC) were left in postmortem ovaries for 30 min (Group I) or 4 h (Group II) at 30°C before aspiration. Progesterone and oestradiol concentrations were measured in the follicular fluid in both groups by immunoassay using an Immulite 2000 analyzer. Immature oocytes were evaluated for MATER and OCT-4 mRNA abundance by real-time PCR (total RNA isolated from pools of 100 oocytes per repeat) or were subjected to in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro culture (IVC). For in vitro embryo production, 455 (Group I) and 470 (Group II) COC were used in three repeats. Progesterone concentration was lower (P ≤ 0.05) in Group II than in Group I. Conversely, oestradiol concentration did not vary between groups. Similarly, Group II oocytes exhibited the highest (P < 0.05) MATER and OCT-4 abundance. For embryo development, there were no significant differences between cleavage rates (72 h post-insemination) between both groups. However, blastocyst (168 h post-insemination) and hatching (216 h post-insemination) rates in Group II were greater (P < 0.05) with 21.3 compared with 30.7% and 54.2 compared with 75.3%, respectively. These results indicate that progesterone concentration in the follicle and the abundance of MATER and OCT-4 transcripts could be good predictors of embryo developmental competence and that retrieving COC 4 h after slaughter could increase blastocyst and hatching rates. This work was supported by COLCIENCIAS COD 122852128473 Colombia.

scholarly journals The developmental competence of bovine oocytes matured in vitro using thymosin beta 4

2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Joanna Romanek ◽  
Joanna Jurkiewicz ◽  
Agnieszka Wierzchoś-Hilczer ◽  
Jolanta Opiela
Keyword(s):  
Developmental Competence ◽  
Vitro Fertilization ◽  
Thymosin Beta 4 ◽  
Maturation Medium ◽  
Significant Difference ◽  
The Mean ◽  
Bovine Oocytes ◽  
Positive Effect

AbstractThe aim of this study was to examine the effect of thymosin beta 4 (Tβ4) during in vitro maturation (IVM) of oocytes and subsequent embryonic development after in vitro fertilization as well as to assess the quality of obtained blastocysts. COCs were matured in vitro in 4 different media: 1. control medium; 2. control media supplemented with 50 ng/mL Tβ4; 3. control media supplemented with 0.5 mg/mL Tβ4; and 4. control media supplemented with 1 mg/mL Tβ4. The quality of the developed blastocysts was analysed by the TUNEL method. The number of cleaved eggs was significantly higher (P < 0.05) when gametes were matured in the presence of 50 ng/mL Tβ4 than it was using the other types of media. Additionally, the largest number of blastocysts was observed when 0.5 mg Tβ4 was added to the medium (P < 0.05). No significant difference was noted in the mean number of apoptotic nuclei per blastocyst or in the mean number of nuclei per blastocyst in any of the analysed groups. In conclusion, Tβ4 supplementation (50 ng/mL) in maturation medium increased the number of cleaved oocytes, and the number of blastocysts obtained increased when 0.5 mg/mL Tβ4 was used. This positive effect was not observed when higher a higher concentration of Tβ4 (1 mg/mL) was used.

scholarly journals Pregnancies and improved early embryonic development with bovine oocytes matured in vitro with 9-cis-retinoic acid

Reproduction ◽  
2003 ◽  
pp. 409-416 ◽  
Author(s):  
CO Hidalgo ◽  
C Diez ◽  
P Duque ◽  
N Facal ◽  
E Gomez
Keyword(s):  
Retinoic Acid ◽  
In Vitro Maturation ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Developmental Potential ◽  
Vitro Fertilization ◽  
Cell Counts ◽  
Bovine Oocytes ◽  
Number Of Cells

Retinoids have an important role in cell growth, morphogenesis and differentiation. In the present study the developmental potential of bovine oocytes was examined after in vitro maturation in the presence of 9-cis-retinoic acid, a vitamin A metabolite, at 5 nmol l(-1) in chemically defined conditions. Experiments studied early in vitro development, blastocyst differential cell counts and the capacity of embryos to establish pregnancy after transfer to recipients. After in vitro fertilization and culture in simple medium, blastocyst development and hatching rates increased in oocytes matured with 9-cis-retinoic acid. Addition of ethanol (used as a solvent for 9-cis-retinoic acid) resulted in higher cell counts and proportions of cells in the inner mass of day 7 blastocysts. Day 8 blastocysts represented most differences observed in the number of cells. In these embryos, ethanol and 9-cis-retinoic acid increased both the number of cells and proportions in the inner mass. However, while ethanol treatment reduced the number of cells in the trophectoderm, 9-cis-retinoic acid did not. The total number of cells was unaffected by treatment within 1 day, although untreated oocytes lead to day 8 blastocysts with reduced total cell counts. Once transferred to recipients, both fresh and vitrified-warmed blastocysts derived from oocytes matured with 9-cis-retinoic acid yielded more pregnancies at day 60. Modifications of retinoid metabolism affect development and trophectoderm differentiation, and in vitro maturation with 9-cis-retinoic acid increased the developmental competence of the oocyte.

Activation of bovine oocytes by protein synthesis inhibitors: new findings on the role of MPF/MAPKs†

2021 ◽  
Author(s):  
Cecilia Valencia ◽  
Felipe Alonso Pérez ◽  
Carola Matus ◽  
Ricardo Felmer ◽  
María Elena Arias
Keyword(s):  
Protein Synthesis ◽  
Kinase Activity ◽  
Cyclin B1 ◽  
Protein Synthesis Inhibitors ◽  
Vitro Fertilization ◽  
New Findings ◽  
Bovine Oocytes ◽  
Dependent Pathway

Abstract The present study evaluated the mechanism by which protein synthesis inhibitors activate bovine oocytes. The aim was to analyze the dynamics of MPF and MAPKs. MII oocytes were activated with ionomycin (Io), ionomycin+anisomycin (ANY) and ionomycin+cycloheximide (CHX) and by in vitro fertilization (IVF). The expression of cyclin B1, p-CDK1, p-ERK1/2, p-JNK, and p-P38 were evaluated by immunodetection and the kinase activity of ERK1/2 was measured by enzyme assay. Evaluations at 1, 4, and 15 hours postactivation (hpa) showed that the expression of cyclin B1 was not modified by the treatments. ANY inactivated MPF by p-CDK1Thr14-Tyr15 at 4 hpa (P &lt; 0.05), CHX increased pre-MPF (p-CDK1Thr161 and p-CDK1Thr14-Tyr15) at 1 hpa and IVF increased p-CDK1Thr14-Tyr15 at 17 hours postfertilization (hpf) (P &lt; 0.05). ANY and CHX reduced the levels of p-ERK1/2 at 4 hpa (P &lt; 0.05) and its activity at 4 and 1 hpa, respectively (P &lt; 0.05). Meanwhile, IVF increased p-ERK1/2 at 6 hpf (P &lt; 0.05); however, its kinase activity decreased at 6 hpf (P &lt; 0.05). p-JNK in ANY, CHX, and IVF oocytes decreased at 4 hpa (P &lt; 0.05). p-P38 was only observed at 1 hpa, with no differences between treatments. In conclusion, activation of bovine oocytes by ANY, CHX, and IVF inactivates MPF by CDK1-dependent specific phosphorylation without cyclin B1 degradation. ANY or CHX promoted this inactivation, which seemed to be more delayed in the physiological activation (IVF). Both inhibitors modulated MPF activity via an ERK1/2-independent pathway, whereas IVF activated the bovine oocytes via an ERK1/2-dependent pathway. Finally, ANY does not activate the JNK and P38 kinase pathways.

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