title: Aromatase-null mice expressing enhanced green fluorescent protein in germ cells provide a model system to assess estrogen-dependent ovulatory responses.
abstract: Enhanced green fluorescent protein (EGFP) has provided us with valuable approaches for tracking living cells. We established a novel line of transgenic mice, which express EGFP in the testis and ovary. Histological analysis demonstrated that spermatids in the testis and oocytes in ovarian follicles beyond preantral stages were positive for EGFP. By exploiting these features, we evaluated ovulatory responses of aromatase-gene (Cyp19a) knockout mouse expressing the EGFP transgene, which is totally anovulatory due to 17β-estradiol (E2) deficiency. Ovulation in the knockout mice was induced by sequential injections of E2 on days 1, 4 and 5, pregnant mare serum gonadotropin on day 4 and human chorionic gonadotropin on day 6. Fluorescent oocytes were readily detectable at 15 h after the last gonadotropin injection in the oviduct under a fluorescence stereomicroscope, even when only one oocyte was present. However, when E2 supplementation on day 4 or day 5 in the regimen was omitted, no ovulated oocytes were detected, indicating that exogenous E2 supplementation at the time of gonadotropin stimulation is necessary to induce ovulation in aromatase-gene knockout mice. Our results further demonstrated that the current mouse line can provide an alternative tool to study germ cell biology, including oogenesis, ovulation and senescence.
authors.affiliation: Department of Biochemistry, School of Medicine, Kochi University, Nankoku, Kochi, 783-8505, Japan, todak@kochi-u.ac.jp.
authors.forename: Katsumi
authors.initials: K
authors.lastname: Toda
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Atsuko
authors.initials: A
authors.lastname: Yamashita
authors.forename: Masaru
authors.initials: M
authors.lastname: Okabe
authors.forename: Masafumi
authors.initials: M
authors.lastname: Ono
authors.forename: Toshiji
authors.initials: T
authors.lastname: Saibara
mesh_terms: D000818:Animals; D001141:Aromatase / Q000172:deficiency* / Q000235:genetics; D015139:Blotting, Southern; D017931:DNA Primers / Q000235:genetics; D004958:Estradiol / Q000008:administration & dosage / Q000494:pharmacology; D005260:Female; D005663:Furans; D005854:Germ Cells / Q000378:metabolism*; D006062:Gonadotropins / Q000008:administration & dosage / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D010060:Ovulation / Q000187:drug effects / Q000502:physiology*; D013876:Thiophenes
chemical_list: C516387:3,4-bis-(2,4,5-trimethyl-thiophen-3-yl)furan-2,5-dione; D017931:DNA Primers; D005663:Furans; D006062:Gonadotropins; D013876:Thiophenes; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004958:Estradiol; D001141:Aromatase