Abstract:
:Retinoic acid (RA), a naturally occurring metabolite of vitamin A, plays an essential role in regulating cellular growth, differentiation, and death in a variety of tissues, particularly during fetal development. However, essentially nothing is known of the effects of RA on fetal gametogenesis. Using a recently validated system of culturing murine fetal ovaries, herein we sought to characterize the actions of RA on female germ cell proliferation and apoptosis during oogenesis. In the absence of trophic hormone support, approximately 90% of the oogonia and oocytes present in fetal ovaries at the start of culture underwent apoptosis over a 72 h culture period (P < 0.05), whereas provision of 0.01-1 microM RA dose dependently maintained germ cell numbers. In fact, ovaries cultured with 0.1 microM RA for 72 h possessed approximately 30% more oogonia and oocytes as compared with the preculture mean number (P < 0.05). Additional experiments, using in situ DNA 3'-end-labeling and cellular morphology to assess apoptosis coupled with 5-bromo-2'-deoxyuridine incorporation to assess proliferation, revealed that RA acts as both a mitogen and a survival factor for female germ cells. Furthermore, the ability of RA to stimulate germ cell proliferation in cultured fetal ovaries was completely suppressed (P < 0.05) by cotreatment with inhibitors of transcription (alpha-amanitin, 0.1 microg/ml) or protein synthesis (cycloheximide, 1.0 microg/ml), whereas RA-mediated suppression of germ cell apoptosis was not affected by cotreatment with either macromolecular synthesis inhibitor (P > 0.05). Moreover, cotreatment of fetal ovaries with 5 microM LY294002, an inhibitor of phosphatidylinositol 3'-kinase, had no effect on RA-promoted germ cell maintenance (P > 0.05). By comparison, the antiapoptotic effects of insulin-like growth factor I on germ cells in cultured fetal ovaries were significantly attenuated by cotreating ovaries with LY294002 (P < 0.05) but not with alpha-amanitin or cycloheximide (P > 0.05). Importantly, the effect of RA on the female germ line was also observed in vivo because a single oral administration of 100 mg/kg RA to timed-pregnant female mice resulted in a significantly (P < 0.05) larger endowment of primordial oocytes in female offspring. That these actions were mediated, at least in part, by specific retinoid receptors was demonstrated by the finding of retinoic acid receptor protein in fetal female gonocytes, as assessed by immunohistochemical localization experiments. Collectively, these data indicate that RA can function, in vitro and in vivo, as a potent germ cell survival factor and mitogen during fetal oogenesis in the mouse.
journal_name
Endocrinologyjournal_title
Endocrinologyauthors
Morita Y,Tilly JLdoi
10.1210/endo.140.6.6826subject
Has Abstractpub_date
1999-06-01 00:00:00pages
2696-703issue
6eissn
0013-7227issn
1945-7170journal_volume
140pub_type
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