Is estradiol mandatory for an adequate follicular and embryo development? A mouse model using aromatase inhibitor (anastrozole).
BACKGROUND
Although high levels of estradiol are found in the follicular fluid, little is known about its necessity for adequate follicular growth, oocyte maturation and embryo development. Arimidex (anastrozole) is a potent aromatase inhibitor capable to induce an in-vivo milieu deprived of estradiol. This study uses a mouse model applying Arimidex to create an in-vivo system lacking of estradiol, in order to explore whether this gonadal steroid hormone is mandatory for folliculogenesis followed by normal fertilization and embryo development.


METHODS
Experiment 1: Immature C57 Black female mice, aged 3-4 weeks were superovulated by 5 IU PMSG given intraperitoneally. A study group (9 mice) was concomitantly injected with 0.1 mg of Arimidex intraperitoneally given the morning day before PMSG, the morning day of PMSG injection and the following two days. The control group (8 mice) was similarly injected with normal saline. Estradiol (E2) and progesterone (P) serum levels were tested 48 hours after PMSG and the ovaries of each mouse blindly examined by a pathologist to evaluate follicular development. Experiment 2: 48 h after PMSG superovulation, hCG (7.5 IU) was injected intraperitoneally, followed by mating. The study group was treated with Arimidex 0.1 mg intraperitoneally daily from a day prior to PMSG injection to the day of sacrifice. The control group was treated similarly by normal saline. Forty-two hours after mating blood was withdrawn for E2 and P levels followed by tubal dissection. Embryos of 2-4 cells were cultured in-vitro and the development to the morula, blastocyst and hatching blastocyst stages were examined 24, 42, and 48 h later.


RESULTS
Experiment 1: A significant reduction of E2 levels was achieved in the Arimidex group in comparison to control group (126.3+/-104.8 and 1910+/-960 pmol/L, respectively; p < 0.0001). Nevertheless, the two groups did not differ by the mean number of follicles (27+/-9.5 and 30.4+/-13.0) or the distribution for antral (65% and 68.4%) and pre-antral (35% and 31.6%) follicles, respectively. Experiment 2: The reduction of estradiol during follicular phase did not hamper follicular development, in-vivo fertilization and in-vitro embryo development. Similar rates of embryo development to the morula stage (90.6% and 86%), blastocyst stage (86% and 89%) and hatching blastocyst (81% and 78%) were achieved in the Arimidex group and the control group, respectively.


CONCLUSIONS
Adequate folliculogenesis is independent of estrogen but is conditioned on gonadotropin stimulation. Moreover, depletion of estradiol in the vicinity of the oocyte did not impair its developmental potential, including its fertilization and development into morulae, blastocysts and hatching blastocysts.
Fatum
Muhammad
M
Department of Obstetrics and Gynecology, Hadassah University Hospital, Ein Kerem, P.O. Box 12000, Jerusalem 91120, Israel. fatum@md.huji.ac.il
Gyo
Yi
Y
Diana
Prus
P
Laufer
Neri
N
Simon
Alex
A
D000077384:Anastrozole; D000818:Animals; D047072:Aromatase Inhibitors / Q000494:pharmacology*; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology*; D004958:Estradiol / Q000502:physiology*; D005260:Female; D006064:Gonadotropins, Equine / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D009570:Nitriles / Q000494:pharmacology*; D006080:Ovarian Follicle / Q000187:drug effects / Q000502:physiology*; D011247:Pregnancy; D013480:Superovulation; D014230:Triazoles / Q000494:pharmacology*
D047072:Aromatase Inhibitors; D006063:Chorionic Gonadotropin; D006064:Gonadotropins, Equine; D009570:Nitriles; D014230:Triazoles; D000077384:Anastrozole; D004958:Estradiol