Effects of epidermal growth factor on early embryonic development after in vitro fertilization of oocytes collected from ewes treated with follicle stimulating hormone

Grazul-Bilska, Anna T.; Choi, Jong Tae; Bilski, Jerzy J.; Weigl, R.M.; Kirsch, James D; Kraft, Kim C.; Reynolds, Lawrence P.; Redmer, Dale A.

doi:10.1016/s0093-691x(02)01192-5

Cited by 36 publications

(24 citation statements)

References 43 publications

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“…Oocytes were cultured in fertilization medium prepared in our laboratory consisting of synthetic oviductal fluid (SOF), 2% heat-inactivated ovine serum collected on day 0–1 of the estrous cycle and 1% penicillin/streptomycin in the presence of capacitated sperm (0.5 to 1 × 10 6 sperm/ml) for 24 h followed by incubation (at 39°C, 5% O 2 , 5% CO 2 and 90% N 2 ) in culture medium until ET (Grazul-Bilska et al 2003, 2006, 2012, 2013). Culture medium consisted of SOF supplemented with bovine serum albumin (BSA; 8 mg/ml; Sigma), glutamine (1 mM), MEM non-essential amino acids (0.01 ml/ml, vol/vol; Sigma), BME amino acids (0.02 ml/ml, vol;/vol; Sigma) and penicillin/streptomycin (100 units/mL penicillin and 100 μg/mL streptomycin).…”

Section: Methodsmentioning

confidence: 99%

Placental development during early pregnancy in sheep: effects of embryo origin on vascularization

Grazul-Bilska¹,

Johnson²,

Borowicz³

et al. 2014

Reproduction

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Utero-placental growth and vascular development are critical for pregnancy establishment that may be altered by various factors including assisted reproductive technologies (ART), nutrition, or others, leading to compromised pregnancy. We hypothesized that placental vascularization and expression of angiogenic factors are altered early in pregnancies after transfer of embryos created using selected ART methods. Pregnancies were achieved through natural mating (NAT), or transfer of embryos from natural mating (NAT-ET), or in vitro fertilization (IVF) or activation (IVA). Placental tissues were collected on day 22 of pregnancy. In maternal caruncles (CAR), vascular cell proliferation was less (P<0.05) for IVA than other groups. Compared to NAT, density of blood vessels was less (P<0.05) for IVF and IVA in fetal membranes (FM), and for NAT-ET, IVF and IVA in CAR. In FM, mRNA expression was decreased (P<0.01–0.08) in NAT-ET, IVF and IVA compared to NAT for vascular endothelial growth factor (VEGF) and its receptor FLT-1, placental growth factor (PGF), neuropilin (NP) 1 and 2, angiopoietin (ANGPT) 1 and 2, endothelial nitric oxide synthase (NOS3), hypoxia inducible factor-1A (HIF1A), fibroblast growth factor (FGF) 2 and its receptor FGFR2. In CAR, mRNA expression was decreased (P<0.01–0.05) in NAT-ET, IVF and IVA compared to NAT for VEGF, FLT-1, PGF, ANGPT1 and TEK. Decreased mRNA expression for 12 of 14 angiogenic factors across FM and CAR in NAT-ET, IVF and IVA pregnancies was associated with reduced placental vascular development, which would lead to poor placental function and compromised fetal and placental growth and development.

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Section: Methodsmentioning

confidence: 99%

Placental development during early pregnancy in sheep: effects of embryo origin on vascularization

Grazul-Bilska¹,

Johnson²,

Borowicz³

et al. 2014

Reproduction

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“…At 24 h after CIDR removal, NAT ewes were exposed to a fertile ram and naturally mated, but for donor ewes from NAT-ET, IVF and IVA groups, estrus was checked twice daily using a vasectomized ram; 5, 86, and 7% of ewes expressed estrus at 24, 36 and 48 h after CIDR removal, respectively. Beginning on Day 13 of the estrous cycle, donor ewes (n=3) for the NAT-ET group were treated twice daily with FSH (Sioux Biochemical, Sioux Center, IA, USA) for 3 d (Day 13, 5 units/injection; Day 14, 4 units/injection and Day 15, 3 units/injection; unit is equivalent to 3.5 g of NIDDK-oFSH-20), whereas donor ewes (n=22) for the IVF and IVA groups were treated with FSH for 2 d (Days 13 and 14, doses as above) following estrus (d 0) as described before [10,39,40]. On Day 15 of the estrous cycle, ewes from the NAT-ET group were exposed to a fertile ram for 24 to 48 h, but for IVF and IVA groups ovaries were collected, and the oocytes were isolated, matured and then fertilized or activated in vitro as previously described in detail [10,36,40,41].…”

Section: Methodsmentioning

confidence: 99%

“…For the IVF and IVA groups, in vitro-generated embryos were transferred on Day 5 after fertilization or activation (Day 1 = day of fertilization or activation) to synchronized recipient ewes (three embryos from the same donor/recipient), as described by Grazul-Bilska et al [10,40]. On Day 22 after mating, fertilization or activation, fetuses and utero-placental tissues were collected from NAT (n=8), NAT-ET (n=7), IVF (n=8), and IVA (n=7) groups.…”

Section: Methodsmentioning

confidence: 99%

Placental development during early pregnancy in sheep: Effects of embryo origin on fetal and placental growth and global methylation

et al. 2013

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The origin of embryos including those created through assisted reproductive technologies (ART) may have profound effects on placental and fetal development, possibly leading to compromised pregnancies associated with poor placental development. To determine the effects of embryo origin on fetal size, and maternal and fetal placental cellular proliferation and global methylation, pregnancies were achieved through natural mating (NAT), or transfer of embryos generated through in vivo (NAT-ET), IVF, or in vitro activation (IVA). On Day 22 of pregnancy, fetuses were measured and placental tissues were collected to immunodetect Ki67 (a marker of proliferating cells) and 5-methyl cytosine (5mC) followed by image analysis, and determination of mRNA expression for three DNA methyltransferases (DNMT). Fetal length and labeling index (proportion of proliferating cells) in maternal caruncles (CAR; maternal placenta) and fetal membranes (FM; fetal placenta) were less (P < 0.001) in NAT-ET, IVF and IVA than in NAT. Expression of 5mC was greater (P < 0.02) in IVF and IVA than in NAT. In CAR, mRNA expression for DNMT1 was greater (P < 0.01) in IVA compared to the other groups, but DNMT3A expression was less (P < 0.04) in NAT-ET and IVA than NAT. In FM, expression of mRNA for DNMT3A was greater (P < 0.01) in IVA compared to the other groups, and was similar in NAT, NAT-ET and IVF groups. Thus, embryo origin may have specific effects on growth and function of ovine utero-placental and fetal tissues through regulation of tissue growth, DNA methylation and likely other mechanisms. These data provide a foundation for determining expression of specific factors regulating placental and fetal tissue growth and function in normal and compromised pregnancies, including those achieved with ART.

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“…The serum-free IVM system reported here demonstrated cleavage rates of 67.7% and blastocyst formation of 34.5% (Table 1). Previous reports have shown that sheep blastocyst production rates can range from 15 to 70% following the IVM of COCs with and without the addition of FCS or oestrus sheep serum (Thompson et al 1989, 1995, Gardner et al 1994, Watson et al 1994, Obrien et al 1996, Walker et al 1996, Wang et al 1998, Grazul-Bilska et al 2003. These serum-based culture systems are commonly supplemented with high pharmacological concentrations of 0.5-10 mg/ml LH and FSH.…”

Section: Discussionmentioning

confidence: 99%

Characterisation of the cellular and molecular responses of ovine oocytes and their supporting somatic cells to pre-ovulatory levels of LH and FSH during in vitro maturation

2012

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The response of Graafian follicles to pre-ovulatory surge levels of FSH and LH in vivo triggers the terminal differentiation of granulosa cells and oocyte maturation. In polyovular species, the LH-driven signalling uses the epidermal growth factor (EGF)-like ligands AREG, EREG and BTC to promote oocyte maturation and cumulus expansion. This experimental series used a physiologically relevant ovine in vitro maturation (IVM) system to evaluate the impact of exposure to pre-ovulatory levels (100 ng/ml) of LH and FSH on ovine cumulus cell expression of EGF-like ligands in vitro. The serum-free sheep IVM system supported high levels (91.4%) of gonadotrophin-induced maturation of cumulus-enclosed oocytes and embryo development to the blastocyst stage (34.5%). Results were equivalent to a serumbased IVM system (85.1% IVM, 25.8% blastocyst rate; PO0.05) but were significantly different (P!0.05) to serum-free medium without gonadotrophins (69.5% IVM; 8.0% blastocyst rate). Ovine BTC was cloned and sequenced. Gonadotrophin-induced AREG, EREG, BTC and EGFR expressions were quantified in cumulus and mural granulosa cells during IVM. A rapid induction of AREG expression was apparent in both cell types within 30 min of gonadotrophin exposure in vitro. LHCGR (LHR) was detected in mural cells and FSHR in both cumulus and mural granulosa cells. The data confirm the involvement of AREG and EGFR during gonadotrophin-induced cumulus expansion, oocyte maturation and the acquisition of developmental competence by sheep oocytes matured in vitro.

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Effects of epidermal growth factor on early embryonic development after in vitro fertilization of oocytes collected from ewes treated with follicle stimulating hormone

Cited by 36 publications

References 43 publications

Placental development during early pregnancy in sheep: effects of embryo origin on vascularization

Placental development during early pregnancy in sheep: effects of embryo origin on vascularization

Placental development during early pregnancy in sheep: Effects of embryo origin on fetal and placental growth and global methylation

Characterisation of the cellular and molecular responses of ovine oocytes and their supporting somatic cells to pre-ovulatory levels of LH and FSH during in vitro maturation

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