Reduction of the developmental competence of sheep oocytes by inhibition of LH pulses during the follicular phase with a GnRH antagonist.
A GnRH antagonist (Antarelix) treatment was used during the breeding season of Romanov ewes, to investigate whether LH pulses are required the day before the preovulatory surge for normal early embryo development in vivo (Expt 1) and in vitro (Expt 2). In Expt 1, at the onset of oestrus after removal of a fluorogestone acetate sponge, group A0.5 (n = 22) received a subcutaneous injection of 0.5 mg Antarelix, and ovulation was induced with an intravenous injection of 3 mg pig LH 24 h later. The control group (group C, n = 20) were untreated. All ewes were mated naturally at 36 and 48 h after oestrus and embryos were recovered 8 days after sponge removal. There were significant differences in the decrease in LH and in the increase in FSH concentration after Antarelix treatment between treated and control groups. The ovulation rate and embryo recovery rate were not significantly different between the two groups but the blastocyst rate was lower (P < 0.0001) in group A0.5 than in group C, with more unfertilized or degenerated oocytes in group A0.5 (69.2%). In Expt 2, 24 h after sponge removal, group A (n = 10) and group B (n = 10) received one subcutaneous injection of 0.5 mg Antarelix. The control group (group C, n = 10) was left untreated. LH pulsatility was re-established in group B with hourly intravenous injections of 5 micrograms ovine LH for 24 h. Oocytes were collected by flushing the oviducts 28 h after the LH surge, and were fertilized and cultured in vitro for 7 days. Ovulation and cleavage rates were not significantly different among the three groups but a higher rate of blastocysts (P < 0.01) was obtained after Antarelix treatment when LH pulsatility was re-established (group B). Oestradiol concentration was strongly depressed (P < 0.0003) after Antarelix treatment in group A, but was maintained after injection of LH pulses in group B, although at a lower value than before the preovulatory surge in the control group. In conclusion, inhibition of endogenous LH pulses 1 day before the preovulatory surge was not essential for ovulation and in vitro fertilization but was associated with a decrease in plasma oestradiol concentrations and inferior embryo development both in vivo and in vitro. When LH pulsatility was re-established, oestradiol concentrations increased and embryo development was restored.
Oussaid
B
B
INRA-Unité Physiologie Reproduction des Mammifères Domestiques, Nouzilly, France.
Mariana
J C
JC
Poulin
N
N
Fontaine
J
J
Lonergan
P
P
Beckers
J F
JF
Cognie
Y
Y
D000704:Analysis of Variance; D000818:Animals; D001755:Blastocyst / Q000166:cytology; D004622:Embryo, Mammalian / Q000166:cytology; D005314:Embryonic and Fetal Development; D004958:Estradiol / Q000097:blood; D005260:Female; D005307:Fertilization in Vitro; D007987:Gonadotropin-Releasing Hormone / Q000037:antagonists & inhibitors*; D007986:Luteinizing Hormone / Q000097:blood / Q000378:metabolism* / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology*; D009865:Oocytes / Q000187:drug effects; D009866:Oogenesis / Q000187:drug effects*; D010062:Ovulation Induction; D011863:Radioimmunoassay; D012756:Sheep / Q000502:physiology*
D009842:Oligopeptides; D007987:Gonadotropin-Releasing Hormone; D004958:Estradiol; D007986:Luteinizing Hormone; C084350:antarelix