Transcript abundance and apoptosis in day-7 porcine blastocyst cultured with exogenous insulin-like growth factor-I

Wasielak, Marta; Fujii, Takashi; Ohsaki, T.; Hashizume, Takumi; Bogacki, Marek; Sawai, Ken

doi:10.1016/j.repbio.2013.01.173

Cited by 7 publications

(5 citation statements)

References 38 publications

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“…However, when these cells had been previously matured in a medium supplemented with IGF-I, the percentage of viable oocytes with intact DNA after vitrification-warming was higher than that observed for oocytes matured and vitrified in standard media. These results match with previous studies conducted with pig blastocysts in which supplementing culture medium with IGF-I was found to reduce DNA fragmentation and apoptosis indexes (Dhali et al, 2011; Ascari et al, 2017) by downregulating the expression of BAX and upregulating that of BCL-2 (Wasielak et al, 2013; Pan et al, 2015). To explain these results, one should bear in mind that BAX is a pro-apoptotic gene involved in cell death and oocyte degeneration.…”

Section: Discussionsupporting

confidence: 91%

“…Two experiments were designed to determine whether the addition of IGF-I to conventional IVM medium and that of GSH to vitrification-warming media improved oocyte survival and development capacity after vitrification and warming. Based on previous reports and preliminary experiments conducted in our lab, the treatments tested were IGF-I at 100 ng·mL −1 (Wasielak et al, 2013) and GSH at 2 mM (0.62 mg·mL −1 ) (Yeste et al, 2014).…”

Section: Methodsmentioning

confidence: 99%

“…In pigs, although IGF-I has been reported to promote the synthesis of hyaluronic acid and the expansion of cumulus cells (Nemcova et al, 2007), its role in IVM remains unclear and has yield inconsistent results. Nevertheless, several studies reported that the addition of IGF-I during in vitro maturation and culture decreases apoptosis in bovine oocytes (Wasielak and Bogacki, 2007; Rodrigues et al, 2016; Ascari et al, 2017) and in porcine blastocysts (Wasielak et al, 2013). Wasielak et al (2013) also demonstrated that adding IGF-1 at a concentration of 100 ng/ml into maturation medium increases the BCL2L1:BAX transcript ratio in pig blastocysts, and the increased expression of anti-apoptotic genes, such as BCL2 , and a decreased expression of apoptotic-related genes, such as BAX , indicates higher blastocyst quality (Chen et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

“…Nevertheless, several studies reported that the addition of IGF-I during in vitro maturation and culture decreases apoptosis in bovine oocytes (Wasielak and Bogacki, 2007; Rodrigues et al, 2016; Ascari et al, 2017) and in porcine blastocysts (Wasielak et al, 2013). Wasielak et al (2013) also demonstrated that adding IGF-1 at a concentration of 100 ng/ml into maturation medium increases the BCL2L1:BAX transcript ratio in pig blastocysts, and the increased expression of anti-apoptotic genes, such as BCL2 , and a decreased expression of apoptotic-related genes, such as BAX , indicates higher blastocyst quality (Chen et al, 2017). Finally, besides reducing apoptosis, IGF-I also enhances the expression of cold-inducible RNA-binding protein (CIRBP), which protects cells from vitrification and warming (Pan et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Supplementing Maturation Medium With Insulin Growth Factor I and Vitrification-Warming Solutions With Reduced Glutathione Enhances Survival Rates and Development Ability of in vitro Matured Vitrified-Warmed Pig Oocytes

et al. 2019

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The present study sought to determine whether in vitro maturation (IVM) of pig oocytes in a medium supplemented with insulin growth factor-I (IGF-I) and subsequent vitrification with or without reduced glutathione (GSH) affect their quality and developmental competence, and the expression of genes involved in antioxidant, apoptotic and stress responses. In Experiment 1, cumulus-oocyte complexes were matured in the absence or presence of IGF-I (100 ng·mL−1) and then vitrified-warmed with or without 2 mM of GSH. Maturation rate was evaluated before vitrification, and oocyte viability, DNA fragmentation and relative transcript abundances of BCL-2-associated X protein (BAX), BCL2-like1 (BCL2L1), heat shock protein 70 (HSPA1A), glutathione peroxidase 1 (GPX1) and superoxide dismutase 1 (SOD1) genes were assessed in fresh and vitrified-warmed oocytes. In Experiment 2, fresh and vitrified-warmed oocytes were in vitro fertilized and their developmental competence determined. Whereas the addition of IGF-I to maturation medium had no effect on oocyte maturation, it caused an increase in the survival rate of vitrified-warmed oocytes. This effect was accompanied by a concomitant augment in the relative transcript abundance of HSPA1A and a decrease of BAX. Furthermore, the addition of GSH to vitrification-warming media increased survival rates at post-warming. Likewise, the action of GSH was concomitant with an increase in the relative abundance of GPX1 and a decrease of BAX transcript. Blastocyst rates of vitrified-warmed oocytes did not differ from their fresh counterparts when IGF-I and GSH were combined. In conclusion, supplementing maturation medium with 100 ng·mL−1 IGF-I and vitrification-warming solutions with 2 mM GSH improves the quality and cryotolerance of IVM pig oocytes, through a mechanism that involves BAX, GPX1 and HSPA1A expression.

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Section: Discussionsupporting

confidence: 91%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Supplementing Maturation Medium With Insulin Growth Factor I and Vitrification-Warming Solutions With Reduced Glutathione Enhances Survival Rates and Development Ability of in vitro Matured Vitrified-Warmed Pig Oocytes

et al. 2019

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“…Decreased total cell count and expression of anti‐apoptotic genes, such as BCL‐2 , and increased expression of apoptotic index genes, including BAX , indicate that the blastocysts are of poor quality. Recent studies have shown that IGF‐1 concentrations of 100 ng/ml increased the BCL‐XL/BAX transcript expression ratio (Wasielak et al., ). Insulin‐like growth factor I also enhanced the yak sperm warming vitality and the cleavage rate of blastocysts (Pan, Cui, Baloch et al., a).…”

Section: Introductionmentioning

confidence: 99%

Insulin‐like growth factor I enhances the developmental competence of yak embryos by modulating aquaporin 3

Chen

Cui

Wen

et al. 2017

Reprod Domestic Animals

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The objective of our present study was to determine the effects of insulin-like growth factor I (IGF-I) on the development of yak (Bos grunniens) embryos after cumulus-oocyte complex (COC) vitrification and warming followed by in vitro fertilization (IVF). In Experiment 1, the yak COCs underwent vitrification and then IVF. Embryos were incubated in synthetic oviductal fluid (SOF) supplemented with four concentrations (0, 50, 100 and 200 ng/ml) of IGF-I, while the yak COCs without vitrification or IGF-I supplementation acted as the control group; the BAX, BCL-2, AQP3mRNA and aquaporin 3 (AQP3) protein expression levels in the five groups of blastocysts were evaluated using quantitative real-time PCR and immunofluorescence analyses. In Experiment 2, the groups described above were fertilized and incubated. The cleavage rate, blastocyst rate, total cell count per blastocyst and the rate of growth of the inner cell mass (ICM) and trophectoderm (TE) were evaluated. The results were as follows: (1) the AQP3 gene expression and protein expression in the control and 100 ng/ml IGF-I treatment groups were the highest. (2) The BAX gene expression was the lowest and the BCL-2 gene expression was the highest in the control and 100 ng/ml IGF-I treatment groups. (3) The rates of cleavage and blastocysts in the control and 100 ng/ml IGF-I groups were higher than those in the other three groups. The total cell count per blastocyst in the vitrified and warmed 100 ng/ml IGF-I group (106.7 ± 4.9) and the control group (107.3 ± 4.2) was higher than that in the vitrified and warmed 0 ng/ml IGF-I (91.2 ± 3.1), 50 ng/ml IGF-I (92.3 ± 3.7) and 200 ng/ml IGF-I (92.4 ± 3.7) groups. Therefore, we conclude that IGF-I can improve yak blastocyst developmental ability, cytomembrane permeability and formation of the blastocyst cavity after COC vitrification by improving the BAX, BCL-2 and AQP3 expression levels.

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Poly(ADP‐ribosyl)ation is involved in pro‐survival autophagy in porcine blastocysts

Lee

Gupta

Kim

et al. 2015

Molecular Reproduction Devel

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Poly(ADP-ribosyl)ation (PARylation) prevents apoptosis through its involvement in pro-survival autophagy in cultured cells; whether or not the same is true for pre-implantation embryos has not yet been documented. In this study, we investigated the participation of PARylation and autophagy in in vitro porcine pre-implantation embryo development. The transcript levels of autophagy-related genes and poly(ADP-ribose) polymerase 1 (PARP1), an enzyme required for PARylation, were transiently up-regulated by fertilization, decreased at the late 1-cell stage, and maintained until the blastocyst stage. LC3, a marker of autophagosomes, and poly(ADP-ribose) (PAR) polymer were present in all stages of pre-implantation development. Exposure of embryos to 3-methyladenine, an autophagy inhibitor, or 3-aminobenzamide, a PARP inhibitor, suppressed the development of blastocysts. Pharmacological inhibition of PARylation further suppressed pro-survival autophagy by decreasing the expression of autophagy-related genes (ATG5, BECLIN1, and LC3) and decreasing LC3 protein abundance while increasing the rate of apoptosis in blastocysts. Deficiency in autophagy also induced abnormal accumulation of SQSTM1/p62 aggregates in porcine blastocysts. Collectively, these data suggest that PARylation is involved in selective autophagic degradation of ubiquitinated proteins, functioning in a pro-survival role, in porcine in vitro-produced embryos. These pro-survival regulatory mechanisms may be important for the control of embryo quality.

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Transcript abundance and apoptosis in day-7 porcine blastocyst cultured with exogenous insulin-like growth factor-I

Cited by 7 publications

References 38 publications

Supplementing Maturation Medium With Insulin Growth Factor I and Vitrification-Warming Solutions With Reduced Glutathione Enhances Survival Rates and Development Ability of in vitro Matured Vitrified-Warmed Pig Oocytes

Supplementing Maturation Medium With Insulin Growth Factor I and Vitrification-Warming Solutions With Reduced Glutathione Enhances Survival Rates and Development Ability of in vitro Matured Vitrified-Warmed Pig Oocytes

Insulin‐like growth factor I enhances the developmental competence of yak embryos by modulating aquaporin 3

Poly(ADP‐ribosyl)ation is involved in pro‐survival autophagy in porcine blastocysts

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