Heat stress during in vitro fertilization decreases fertilization success by disrupting anti‐polyspermy systems of the oocytes

Sakatani, Miki; Yamanaka, Kenichi; Balboula, Ahmed Z.; Takenouchi, Naoki; Takahashi, Masashi

doi:10.1002/mrd.22441

Cited by 33 publications

(32 citation statements)

References 61 publications

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“…Intracellular ROS were immediately measured using fluorescent probes as previously described (Sakatani, Yamanaka, Balboula, Takenouchi, & Takahashi, ). Briefly, embryos were placed in PVP‐PBS containing 10 µM 2′, 7′‐dichlorodihydrofluorescein diacetate (DCHFDA; Molecular Probes, Eugene, OR) for 15 min at 38.5°C in a humidified atmosphere containing 5% CO 2 in the air and washed twice with PVP‐PBS.…”

Section: Methodsmentioning

confidence: 99%

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Sericin enhances the developmental competence of heat‐stressed bovine embryos

Khatun

Egashira

Sakatani

et al. 2018

Molecular Reproduction Devel

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We investigated the effects of sericin on the developmental competence of bovine embryos exposed to heat stress (HS). Putative zygotes were cultured with sericin and subjected to HS (40.5°C for 6 hr) on Day 2 or 7 followed by continuous culture at 38.5°C until Day 8. Day 2 HS significantly decreased blastocyst development on Day 8 as well as mitochondrial activity, and significantly increased the amount of intracellular reactive oxygen species and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL)-positive cells, whereas Day 7 HS only significantly decreased mitochondrial activity and increased the number of TUNEL-positive cells in Day 8 blastocysts. These detrimental effects were neutralized by sericin supplementation. Next, to investigate the potential production of blastocysts with high viability in terms of thermotolerance, embryos were cultured with sericin until Day 7, and then exposed to HS in the sericin-free medium. TUNEL-positive cell numbers were significantly lower in blastocysts produced by sericin culture than in control blastocysts. Transcript abundance for HSPA1A and BAX was significantly decreased but IFNT2 levels were increased in blastocysts produced by sericin culture. In conclusion, these findings demonstrate the anti-oxidative and anti-apoptotic activities of sericin, and the potential use of sericin to produce embryos with high viability in vitro.

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Section: Methodsmentioning

confidence: 99%

“…Intracellular ROS were immediately measured using fluorescent probes as previously described (Sakatani, Yamanaka, Balboula, Takenouchi, & Takahashi, 2015). Briefly, embryos were placed in PVP-PBS containing…”

Section: Detection Of Oxidative Stress In Embryosmentioning

confidence: 99%

Sericin enhances the developmental competence of heat‐stressed bovine embryos

Khatun

Egashira

Sakatani

et al. 2018

Molecular Reproduction Devel

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“…The in vitro maturation, fertilization, and culture procedures used in this study were performed as described previously (Sakatani, Yamanaka, Balboula, Takenouchi, & Takahashi, ). Briefly, ovaries were obtained from Japanese Black cows slaughtered at a local abattoir.…”

Section: Methodsmentioning

confidence: 99%

Normal DNA methylation status in sperm from a somatic cell cloned bull and their fertilized embryos

Yamanaka

Yamashita

Khatun

et al. 2018

Animal Science Journal

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Epigenetic reprogramming confers totipotency even during somatic cell nuclear transfer (SCNT), which has been used to clone various animal species. However, as even apparently healthy cloned animals sometimes have aberrant epigenetic status, the harmful effects of these defects could be passed onto their offspring. This is one of the biggest obstacles for the application of cloned animals for livestock production. Here, we investigated the DNA methylation status of four developmentally regulated genes (PEG3, XIST, OCT4, and NANOG) in sperms from a cloned and a non-cloned bull, and blastocysts obtained by in vitro fertilization using those sperms and SCNT. We found no differences in the methylation status of the above genes between cloned and non-cloned bull sperms. Moreover, the methylation status was also similar in blastocysts obtained with cloned and non-cloned bull sperms. In contrast, the methylation status was compromised in the SCNT blastocysts. These results indicate that sperm from cloned bulls would be adequately reprogrammed during spermatogenesis and, thus, could be used to produce epigenetically normal embryos. This study highlights the normality of cloned bull offspring and supports the application of cloned cattle for calf production.

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“…However, experiment work of Sakatani et al (2015), revealed that embryos that reached the blastocyst stage which were exposed to heat stress at the early stages of the oocyte does not affect the total cell number of the trophectoderm cell and the inner cell mass ratio any differently compared to the none heat shocked embryos during in vitro culture. Therefore, all these findings suggest that embryos that have passed through the effects of heat exposure might results in the reduction of cell nuclei number or restore a normal viability of nuclei cell number regardless of exposure to heat stress (Sakatani et al, 2004).…”

Section: Nuclei Cell Analysis Of Produced Blastocystmentioning

confidence: 98%

145 the Role of Cell Apoptosis on in Vitro-Produced Beef Cattle Embryos

Nkadimeng

2017

Reprod. Fertil. Dev.

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Successful in vitro development of embryos is dependent upon maintenance of cellular function in the embryonic microenvironment. Exposure of pre-implantation embryos to a variety of cellular stresses can induce apoptosis in all or a fraction of blastomeres. Among the conditions that can induce apoptosis in cattle embryo production in vitro are heat shock, media composition, oxygen levels, and tumour necrosis factor (TNF)-α. The aim of the study was therefore to evaluate the in vitro culture environment of embryos in terms of cell apoptosis through caspase and DNA fragmentation evaluation. A total of 1200 immature oocytes were collected at slaughter from indigenous South African cow ovaries. The cumulus-oocyte complexes were randomly allocated into 2 maturation temperatures, 39°C and 41°C (200/temperature /replicated 6 times), and cultured in M199 + FSH-LH-oestradiol medium under oil at 100% humidity and 5% CO2 for 24 h. Postmaturation, oocytes were subjected to normal subsequent embryo conditions (Rynkowska et al. 2011 Biotechnol. Comput. Biol. Bionanotechnol. 92, 45–53). All matured oocytes were fertilized for 6 h with frozen-thawed Nguni bull semen from 10 bulls replicated 5 times at a concentration of 265 × 106 sperm cells/mL in Bracket and Oliphant medium under oil. The presumptive zygotes from each treatment were cultured into SOF-BSA medium under oil and incubated at 39°C for assessment of cleavage rate post IVF. Produced embryos were divided into 3 groups: 2–4 cell, ≥8 cell embryos, and blastocyst. Two- to four-cell embryos were removed at Day 2, ≥8 cell at Day 5 (mainly 8-cell, morula, and cavitated blastocyst), and expanded blastocyst at Day 7 of embryo culture to determine caspase activity and DNA fragmentation for evidence of apoptosis. Caspase was performed using colourimetric assay on a 96-well microplate reader and monitored at 450 nm reference filter. The DNA fragmentation was examined using the TUNEL assay method and imaging was done using the Alexa Fluor® 488 emission at 519 nm. Data were analysed using SAS 9.2 software (SAS Institute Inc., Cary, NC, USA), and Shapiro-Wilks test was used on the standardized residuals to test for deviations from normality, means of significant effects were compared using Student’s t-l.s.d. at the 95% confidence interval. There was no significant difference on the cleavage rates [39°C (72.0 ± 22.70) and 41°C (67.2 ± 18.9)] and blastocysts [39°C (11.4 ± 2.6) and 41°C (11.2 ± 6.3)] from both maturation temperatures. Caspase activity showed no significant difference on 2–4 cell [39°C (0.015 ± 0.001) and 41°C (0.016 ± 0.002)] and ≥8 cell embryos at both temperatures [39°C (0.022 ± 0.007a) and 41°C (0.032 ± 0.013)]. However, blastocyst differed significantly at both temperatures [39°C (0.037 ± 0.012) and 41°C (0.053 ± 0.005)]. A higher (P < 0.05) DNA fragmentation was observed at 2–4 cell (6.5 ± 2.9), ≥8 cell (13.3 ± 3.1), and blastocyst (48.0 ± 8.2) for embryos produced from 41°C matured oocytes compared to the 39°C maturation group [2–4 cell (2.2 ± 1.2), ≥8 cell (4.5 ± 1.9), and blastocyst (9.7 ± 6.7)]. It is therefore concluded that embryos produced from 41°C matured oocytes can have the same developmental capacity as the 39°C maturation group. Moreover, both temperatures can show signs of apoptosis, however, with more apoptosis evidence in the 41°C than the 39°C maturation group.

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Heat stress during in vitro fertilization decreases fertilization success by disrupting anti‐polyspermy systems of the oocytes

Cited by 33 publications

References 61 publications

Sericin enhances the developmental competence of heat‐stressed bovine embryos

Sericin enhances the developmental competence of heat‐stressed bovine embryos

Normal DNA methylation status in sperm from a somatic cell cloned bull and their fertilized embryos

145 the Role of Cell Apoptosis on in Vitro-Produced Beef Cattle Embryos

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