Role of FOXO1A in the regulation of insulin-like growth factor-binding protein-1 in human endometrial cells: interaction with progesterone receptor.

Insulin-like growth factor-binding protein-1 (IGFBP1) is a major secretory product of the decidualized endometrium. In the present study, we investigated the role of two transcription factors, progesterone receptor (PGR) and a member of the forkhead box class O family of transcription factors (FOXO1A), in the regulation of the IGFBP1 gene in endometrial cells. Human endometrial fibroblasts (HuF) expressed FOXO1A, progesterone receptor A (PGRA), and progesterone receptor B (PGRB) proteins, whereas the endometrial adenocarcinoma cell line, HEC-1B cells, expressed only FOXO1A and no detectable PGR proteins. When FOXO1A expression was silenced using small interference RNA, IGFBP1 expression decreased in both HuF and HEC-1B cells. Using the chromatin immunoprecipitation technique, we demonstrated that liganded PGR was recruited to the IGFBP1 promoter region (-358 to -49). In addition, immunoprecipitation of HuF nuclear proteins with a PGR antibody followed by immunoblotting with anti-FOXO1A revealed that these two proteins interact in these cells. Reporter studies demonstrated that whereas liganded PGRA or PGRB increased a progesterone response element-linked reporter construct, pPRE/ GRE.E1b.Luc, coexpression of FOXO1A inhibited the PGRB response in HuF and synergistically increased PGRA and PGRB response in HEC-1B cells. Furthermore, in HEC-1B cells, FOXO1A increased IGFBP1 promoter activity, and coexpression of PGRA or PGRB further increased the promoter activity in a cooperative manner. In HuF, the response to FOXO1A and PGR was not additive; in fact, it was lower than the sum of the individual responses. Thus, FOXO1A and PGR associate with one another, and each influences the transactivating potential of the other. The cell type-dependent responses strongly implicate the involvement of other cofactors.

Embryo implantation during the short luteal phase of the corn mouse, Calomys musculinus, and the apparent lack of a lactational diapause in South American murid rodents.

As the corn mouse, Calomys musculinus, has a short luteal phase (2-3 days) that is not prolonged after copulation, it was hypothesized that (i) implantation would occur at the end of this phase, that is, earlier than it occurs in most murid species that have been studied, and (ii) a lactational embryonic diapause would not occur during the luteal phase. These hypotheses were tested in females that had copulated during postpartum oestrus and were either lactating or not lactating. Data were recorded from day 3 to day 5 of pregnancy (day 1 = day after coitus), at both 03:00-05:00 h and 17:00-19:00 h. Evidence of implantation in both non-lactating and lactating animals was apparent at 03:00-05:00 h on day 4 (endometrial 'blue reaction' in all cases and failure to recover free uterine embryos in some cases) and implantation swellings appeared within 24 h in both groups. In another experiment, the increase in duration of interbirth intervals in continuously mated females and their correlation with the number of suckling young were compared among C. musculinus, C. laucha, Akodon molinae (South American murid species) and Peromyscus maniculatus (a North American murid in which a lactational embryonic diapause has been shown). The results were indicative of a lactational embryonic diapause in the North American species, but not in the South American species. It was concluded that in C. musculinus (i) implantation occurs at the end of the spontaneous luteal phase, and (ii) that a lactational embryonic diapause does not occur: the absence of a lactational embryonic diapause may be common to other South American murid rodents.

Evidence that a novel type of progestational phase control occurs in the corn mouse, a South American murid rodent.

Most Muridae display a short luteal phase that becomes functional as a consequence of the prolactin release induced by the stimulation of copulation and/or lactation. The corn mouse also shows a short luteal phase, and we wanted to know whether copulation and/or lactation would release prolactin and maintain progesterone secretion in this species. Females in postpartum estrus were either allowed to copulate with an intact male or not, and either to lactate their young or not. Afterward, plasma progesterone was elevated over the baseline level only in females that had copulated and were bearing growing embryos (whether or not they were lactating), while prolactin was elevated only in lactating females. In another experiment, endometrial scratching induced decidualization both in females that had copulated with a vasectomized male and in those that had not copulated; sham operations had no effect in either case. Progesterone levels were elevated in decidualized animals as compared with their sham-operated controls. Results indicate that the initiation of the progestational phase in the corn mouse is not dependent on prolactin release. A short luteal phase during which nidation may occur has not yet been described in any other mammal.