DOI: 10.5061/dryad.bvq83bk69
title: Data from: Forkhead box a2 (FOXA2) impacts uterine decidualization and fetoplacental development

description: Glands of the uterus are essential for the establishment of pregnancy in
 mice and likely humans. Forkhead box a2 (FOXA2) is a transcription factor
 expressed specifically in the glands of the uterus and a critical
 regulator of uterine gland differentiation, development and function. Mice
 with a conditional deletion of FOXA2 in the adult uterus, created using
 the lactotransferrin iCre (Ltf-iCre) model, have a morphologically normal
 uterus with glands, but lack a considerable number of FOXA2-dependent
 GE-expressed genes. Adult FOXA2 conditional knockout (cKO; Ltf-iCre:Foxa2
 flox) mice are infertile due to defective embryo implantation arising from
 a lack of leukemia inhibitory factor (LIF), a critical implantation factor
 of uterine gland origin. Intraperitoneal injections of LIF can initiate
 embryo implantation in the uterus of adult FOXA2 cKO mice with pregnancies
 maintained to term. Here, we tested the hypothesis that FOXA2-regulated
 genes in the uterine glands impact development of the decidua, placenta
 and fetus using a littermate study design. On gestational day (GD) 8.5,
 the anti-mesometrial and mesometrial decidua transcriptome was noticeably
 altered in FOXA2 cKO mice. Viable fetuses were reduced in FOXA2 cKO mice
 on GDs 12.5 and 17.5. Sex-dependent differences in fetal weight, placenta
 histoarchitecture, and the placenta and metrial gland transcriptome were
 observed between control and FOXA2 cKO mice. The transcriptome of the
 placenta with a female fetus was considerably more altered than the
 placenta with a male fetus in FOXA2 cKO dams. These studies reveal a
 previously unrecognized biological role for FOXA2 and uterine glands on
 fetoplacental development that exhibits sexual dimorphism. Thus, uterine
 glands and, by inference, their products program fetoplacental development
 with potential sexually dimorphic impacts on offspring health into
 adulthood.
description: FOXA2 was conditionally deleted in the uterus. Floxed Foxa2 mice
 were crossed with or LtfiCre mice to generate conditional knockout
 animals. Floxed Foxa2f/f mice (stock no. 022620) and LtfiCre mice (stock
 no. 026030) were obtained from The Jackson Laboratory. Gestational time
 points were obtained by the mating of 8- to 10-wk old Control (Foxa2f/f)
 and FOXA2 cKO (LtfiCr/+Foxa2f/f) females to wild-type CD-1 male mice with
 the day that a vaginal plug was observed was considered day 0.5 of
 gestation. For FOXA2 cKO dams, mice received i.p. injections of
 recombinant mouse LIF (10 μg in saline; catalog #554008, BioLegend) at
 1000 h and 1800 h on GD 3.5. Freshly collected gestation day 8.5
 mesometrial and anti-mesometrial decudua from littermate  control and
 Foxa2 conditional knockout uterus were snap frozen in liquid nitrogen.
 Then on gestation day 12.5, placenta and metrial gland from littermate
  control and Foxa2 conditional knockout uterus were snap frozen in liquid
 nitrogen.  Also, freshly collected gestation day 17.5 placenta and metrial
 gland from littermate  control and Foxa2 conditional knockout uterus were
 snap frozen in liquid nitrogen. Total RNA was isolated from frozen
 speciment using a standard TRIzol-based protocol (Catalog 15596026, Thermo
 Fisher). To eliminate genomic DNA contamination, extracted RNA was treated
 with DNase I and purified using a RNeasy MinElute Cleanup Kit (Qiagen).
 The quality and concentration of RNA were determined using a Fragment
 Analyzer (Advanced Analytical Technologies).  RNA-seq was peformed to
 profile gene expresion.