Transition of Cleavage Divisions During In Vitro Development of Bovine Embryos

Ushijima, Hitoshi; Akiyama, Kiyoshi; Tajima, Toshio

doi:10.1274/jmor.26.42

Cited by 3 publications

(2 citation statements)

References 26 publications

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“…A representative image of embryos acquired 8 days after fertilization can be found on Figure 10 . A comprehensive visual identification of bovine embryo developmental stages can be found in Wei et al (2017) and Ushijima and Akiyama (2009) .…”

Section: Resultsmentioning

confidence: 99%

Bovine In Vitro Oocyte Maturation and Embryo Production Used as a Model for Testing Endocrine Disrupting Chemicals Eliciting Female Reproductive Toxicity With Diethylstilbestrol as a Showcase Compound

et al. 2022

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Endocrine disrupting chemicals (EDCs) can interfere with normal hormonal action and regulation. Exposure of women to EDCs has been associated with adverse reproductive health outcomes. The assays currently used to identify EDCs that elicit female reproductive toxicity lack screening tests that address effects on the maturation of oocytes, a process that enables them to be fertilized and develop into embryos. Here, a screening method employing the bovine model of in vitro oocyte maturation and embryo production is described. Endpoints explored address important events in oocyte maturation and developmental competence acquisition. To test the method, the effects of the known human EDC diethylstilbestrol (DES; an estrogen receptor agonist) were evaluated in a range of concentrations (10–9 M, 10–7 M, 10–5 M). Bovine oocytes were exposed to DES during in vitro maturation (IVM) or embryos were exposed during in vitro embryo culture (IVC). The endpoints evaluated included nuclear maturation, mitochondrial redistribution, cumulus cell expansion, apoptosis, and steroidogenesis. DES-exposed oocytes were fertilized to record embryo cleavage and blastocyst rates to uncover effects on developmental competence. Similarly, the development of embryos exposed to DES during IVC was monitored to assess the impact on early embryo development. Exposure to 10–9 M or 10–7 M DES did not affect the endpoints addressing oocyte maturation or embryo development. However, there were considerable detrimental effects observed in oocytes exposed to 10–5 M DES. Specifically, compared to vehicle-treated oocytes, there was a statistically significant reduction in nuclear maturation (3% vs 84%), cumulus expansion (2.8-fold vs 3.6-fold) and blastocyst rate (3% vs 32%). Additionally, progesterone and pregnenolone concentrations measured in IVM culture media were increased. The screening method described here shows that bovine oocytes were sensitive to the action of this particular chemical (i.e., DES), albeit at high concentrations. In principle, this method provides a valuable tool to assess the oocyte maturation process and early embryo development that can be used for reproductive toxicity screening and possibly EDC identification. Further studies should include EDCs with different mechanisms of action and additional endpoints to further demonstrate the applicability of the bovine oocyte model for chemical risk assessment purposes and EDC identification.

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Section: Resultsmentioning

confidence: 99%

Bovine In Vitro Oocyte Maturation and Embryo Production Used as a Model for Testing Endocrine Disrupting Chemicals Eliciting Female Reproductive Toxicity With Diethylstilbestrol as a Showcase Compound

et al. 2022

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“…La celularidad es considerada como uno de los indicadores más importantes de calidad embrionaria (Ushijima et al, 2009). En este estudio se encontró el mayor número de células (p<0.05) por embrión en T4 (145.6) en comparación con los demás grupos experimentales (Cuadro 2).…”

Section: Discussionunclassified

Efecto del suplemento Insulina-Transferrina-Selenio (ITS) sobre la tasa de desarrollo y calidad de embriones bovinos in vitro

Acosta Lobo,

Robledo Salgado,

Londoño Franco

et al. 2024

Rev. investig. vet. Perú

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The aim of this study was to evaluate the effect of Insulin-Transferrin-Selenium (ITS) supplementation in embryo culture on the blastocyst rate and embryo quality. The bovine zygotes obtained by in vitro maturation (MIV) and fertilization (IVF) were transferred to Evolve medium in 4 groups (T1: Without ITS supplement; T2: ITS the first 48 h of culture (CIV1): T3: ITS after day 2 of culture (CIV2), T4: ITS during all culture time (CIV1 + CIV2). All procedures were performed at 38.5 °C, 5% CO2 and 90% relative humidity. The cleavage and blastocyst rates were determined at 48 h and day 8 of culture. The total cell number and the production of reactive oxygen species (ROS) in the expanded blastocysts were determined by staining the nucleus with Hoechst and the oxidation of the fluorescent probe dichlorodihydrofluorescein diacetate (H2DCFDA). The T4 cultured embryos presented the highest rate of blastocysts (32.6%) and total cell number (145.6) compared to the control group without STI (26.2% and 98.6, respectively; p<0.05). Likewise, the production of ROS in T4 blastocysts was lower (p<0.05) than that obtained in the other study groups. The results show that the addition of the ITS supplement throughout the embryo culture time has beneficial effects on the percentage of blastocysts and embryo quality, assessed by the number of total cells and the production of ROS.

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Protocol for the electroporation of CRISPR-Cas for DNA and RNA targeting in Bos taurus zygotes

Biase,

Schettini

2024

STAR Protocols

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Transition of Cleavage Divisions During In Vitro Development of Bovine Embryos

Cited by 3 publications

References 26 publications

Bovine In Vitro Oocyte Maturation and Embryo Production Used as a Model for Testing Endocrine Disrupting Chemicals Eliciting Female Reproductive Toxicity With Diethylstilbestrol as a Showcase Compound

Bovine In Vitro Oocyte Maturation and Embryo Production Used as a Model for Testing Endocrine Disrupting Chemicals Eliciting Female Reproductive Toxicity With Diethylstilbestrol as a Showcase Compound

Efecto del suplemento Insulina-Transferrina-Selenio (ITS) sobre la tasa de desarrollo y calidad de embriones bovinos in vitro

Protocol for the electroporation of CRISPR-Cas for DNA and RNA targeting in Bos taurus zygotes

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