TY - JOUR
T1 - Amphiregulin promotes the maturation of oocytes isolated from the small antral follicles of the rhesus macaque
AU - Peluffo, Marina C.
AU - Ting, Alison Y.
AU - Zamah, Alberuni M.
AU - Conti, Marco
AU - Stouffer, Richard L.
AU - Zelinski, Mary B.
AU - Hennebold, Jon D.
N1 - Funding Information:
This research was supported by the Oncofertility Consortium NIH 1 UL1 RR024926 (R01-HD058294, PL1-EB008542), the NIH/Eunice Kennedy Shriver NICHD Specialized Cooperative Centers Program in Reproduction and Infertility Research (U54 HD18185), the NIH/ Eunice Kennedy Shriver NICHD Contraceptive Development and Research Center (U54 HD55744), NIH/NCRR-RR000163, NIH R01 HD052909 (MC) and a Lalor Foundation Postdoctoral Basic Research Fellowship (MCP). The use of the Leica confocal was supported by grant number S10RR024585. The statistical analyses of the data from the 27 experiments regarding oocyte maturation was performed by the statistician Byung Park with support from the ONPRC Biostatistics Unit, grant number RR000163 from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH).
PY - 2012/8
Y1 - 2012/8
N2 - background: In non-primates, the epidermal growth factor (EGF) and EGF-related ligands such as amphiregulin (AREG) serve as critical intermediates between the theca/mural cells and the cumulus-oocyte-complex (COC) following the mid-cycle LH surge. Studies were designed in primates (1) to analyze AREG levels in follicular fluid (follicular fluid) obtained from pre-ovulatory follicles, as well as (2) to assess dose-dependent effects of AREG on oocytes from small antral follicles (SAFs) during culture, including meiotic and cytoplasmic maturation. methods: Controlled ovulation protocols were performed on rhesus monkeys (n = 12) to determine AREG content within the single, naturally selected dominant follicle after an ovulatory stimulus. Using healthy COCs (n = 271) obtained from SAFs during spontaneous cycles (n = 27), in vitro maturation (IVM) was performed in the absence or presence of physiological concentrations of AREG (10 or 100 ng/ml) with or without gonadotrophins (FSH, 75 mIU/ml; LH, 75 mIU/ml). At the end of the culture period, oocyte meiotic maturation was evaluated and ICSI was performed (n = 111), from which fertilization and early embryo development was followed in vitro. results: AREG levels in follicular fluid from pre-ovulatory follicles increased (P < 0.05) following an ovulatory bolus of hCG at 12, 24 and 36 h post-treatment. At 12 h post-hCG, AREG levels in follicular fluid ranged from 4.8 to 121.4 ng/ml. Rhesus macaque COCs incubated with 10 ng/ml AREG in the presence of gonadotrophins displayed an increased percentage of oocytes that progressed to the metaphase II (MII) stage of meiosis (82 versus 56%, P < 0.05) and a decreased percentage of metaphase I (MI) oocytes (2 versus 23%, P , 0.05) relative to controls, respectively. The percentage of either MI or MII oocytes at the end of the culture period was not different between oocytes cultured with 100 ng/ml AREG or in media alone. Fertilization and first cleavage rates obtained by ICSI of all IVM MII oocytes were 93 and 98%, respectively, and did not vary among treatment groups. Of the MII oocytes that fertilized (n = 103), 37 were randomly selected and maintained in culture to assess developmental potential. A total of 13 early blastocysts were obtained, with four embryos developing to expanded blastocysts. conclusions: These data indicate that AREG levels increase in rhesus macaque pre-ovulatory follicles after an ovulatory stimulus, and a specific concentration of AREG (10 ng/ml) enhances rhesus macaque oocyte nuclear maturation but not cytoplasmic maturation from SAFs obtained during the natural menstrual cycle. However, owing to the small number of samples in some treatment groups, further studies are now required.
AB - background: In non-primates, the epidermal growth factor (EGF) and EGF-related ligands such as amphiregulin (AREG) serve as critical intermediates between the theca/mural cells and the cumulus-oocyte-complex (COC) following the mid-cycle LH surge. Studies were designed in primates (1) to analyze AREG levels in follicular fluid (follicular fluid) obtained from pre-ovulatory follicles, as well as (2) to assess dose-dependent effects of AREG on oocytes from small antral follicles (SAFs) during culture, including meiotic and cytoplasmic maturation. methods: Controlled ovulation protocols were performed on rhesus monkeys (n = 12) to determine AREG content within the single, naturally selected dominant follicle after an ovulatory stimulus. Using healthy COCs (n = 271) obtained from SAFs during spontaneous cycles (n = 27), in vitro maturation (IVM) was performed in the absence or presence of physiological concentrations of AREG (10 or 100 ng/ml) with or without gonadotrophins (FSH, 75 mIU/ml; LH, 75 mIU/ml). At the end of the culture period, oocyte meiotic maturation was evaluated and ICSI was performed (n = 111), from which fertilization and early embryo development was followed in vitro. results: AREG levels in follicular fluid from pre-ovulatory follicles increased (P < 0.05) following an ovulatory bolus of hCG at 12, 24 and 36 h post-treatment. At 12 h post-hCG, AREG levels in follicular fluid ranged from 4.8 to 121.4 ng/ml. Rhesus macaque COCs incubated with 10 ng/ml AREG in the presence of gonadotrophins displayed an increased percentage of oocytes that progressed to the metaphase II (MII) stage of meiosis (82 versus 56%, P < 0.05) and a decreased percentage of metaphase I (MI) oocytes (2 versus 23%, P , 0.05) relative to controls, respectively. The percentage of either MI or MII oocytes at the end of the culture period was not different between oocytes cultured with 100 ng/ml AREG or in media alone. Fertilization and first cleavage rates obtained by ICSI of all IVM MII oocytes were 93 and 98%, respectively, and did not vary among treatment groups. Of the MII oocytes that fertilized (n = 103), 37 were randomly selected and maintained in culture to assess developmental potential. A total of 13 early blastocysts were obtained, with four embryos developing to expanded blastocysts. conclusions: These data indicate that AREG levels increase in rhesus macaque pre-ovulatory follicles after an ovulatory stimulus, and a specific concentration of AREG (10 ng/ml) enhances rhesus macaque oocyte nuclear maturation but not cytoplasmic maturation from SAFs obtained during the natural menstrual cycle. However, owing to the small number of samples in some treatment groups, further studies are now required.
KW - Amphiregulin
KW - Embryo development
KW - Oocyte maturation
KW - Rhesus monkeys
KW - Small antral follicle
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U2 - 10.1093/humrep/des158
DO - 10.1093/humrep/des158
M3 - Article
C2 - 22593432
AN - SCOPUS:84867805022
SN - 0268-1161
VL - 27
SP - 2430
EP - 2437
JO - Human Reproduction
JF - Human Reproduction
IS - 8
ER -