GATA2 is expressed at critical times in the mouse uterus during pregnancy

doi:10.1016/j.gep.2012.03.004

Gene Expression Patterns

Volume 12, Issues 5–6, May–June 2012, Pages 196-203

https://doi.org/10.1016/j.gep.2012.03.004 Get rights and content

Abstract

In mammals, such as mouse and human, timely production of the progesterone receptor (PR) in the proper uterine compartments is critical for preparing the uterus for the initiation and maintenance of pregnancy. Developmentally, the expression of GATA2, a member of the six member zinc-finger family of transcription factors, has been shown to be necessary for multiple non-related tissues, such as the hematopoietic system, adipose maturation and the urogential system. We recently identified Gata2 as a potential progesterone target gene in the mouse uterus; however, the expression of the GATA genes in the mouse uterus during pregnancy has not been demonstrated. In the present study, we examined the expression of GATA2 protein during the phases of pregnancy, including early pregnancy where progesterone (P4) signaling is critical in order to facilitate the window of receptivity for embryo implantation and during the decidualization of the uterine stroma, a process of cellular proliferation and differentiation which is necessary for maintenance of the invading embryo until placentation occurs. Here, we report that GATA2 protein is expressed in the uterine luminal and glandular epithelium pre-implantation, spatio-temporally co-localizing with that of the PR. Additionally, GATA2 continues to be expressed in the decidualized stroma throughout early pregnancy indicating a role in the maintenance of decidual cells. Based on these findings, we conclude that GATA2 is expressed during critical phases of early pregnancy, similar to that of the PR, and that it may play a major role in mediating P4 signaling in the mouse uterus.

Highlights

► We confirm GATA2 as a P4 responsive target in the uterus. ► We examined GATA2 protein expression during early pregnancy and decidualization. ► GATA2 peak expression in early pregnancy co-localizes with the progesterone receptor. ► Expression of GATA2 persists through embryo invasion of the stroma in surrounding decidual cells.

Section snippets

Confirmation of Gata2 as a progesterone target gene in the murine uterus

To verify Gata2 as a uterine P4 target gene we utilized both in vitro and in vivo methods. Ligand induced nuclear receptor regulation of gene expression, including that of the PR, acts by translocation into the nucleus and interacting with the chromatin through the receptors’ DNA binding domain and subsequent recruitment of co-regulators (Li and O’Malley, 2003). We utilized in vitro cell culture to investigate the ability of the PR to regulate expression of an approximately 1 kb region of the

Transfection and luciferase assay

Human endometrial epithelial cells (HEC-1A) were transfected according to manufacturer’s instructions using Qiagen SuperFect reagent (Qiagen, Valencia, CA, USA) with 200 ng of ∼900 bp region of the mouse Gata2 proximal promoter fused to the pGL3 luciferase vector (Promega, Madison, WI) along with 50 ng of human PR-A, human PR-B, or pcDNA empty vector control. The cells were treated with 10⁻⁸ M R5020 (Sigma, St. Louis, MO, USA) or vehicle control (ethanol) in McCoy’s 5A media (Invitrogen, Carlsbad,

Acknowledgements

The authors are thankful to Yan Ying, M.S. for development of the Immunofluorescence protocol and the personnel of the Genetically Engineered Mouse Core of Baylor College of Medicine. This project was supported by NIH Grant R01HD042311 (to F.J.D.), NIH Grant R01HD057873 (to J.W.J.), NIH518 Grant R01CA77530 (to J.P.L).

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Dysregulated expression of GATA2 and GATA6 transcription factors in adenomyosis: implications for impaired endometrial receptivity
2024, F and S Science
To study the effect of adenomyosis on the localized expression of the GATA binding proteins 2 and 6 (GATA2 and GATA6) zinc-finger transcription factors that are involved in proliferation of hematopoietic and endocrine cell lineages, cell differentiation, and organogenesis, potentially leading to impaired endometrial implantation.
Laboratory based experimental study.
Academic hospital and laboratory.
Human endometrial stromal cells (HESCs) of reproductive age patients, 18–45 years of age, with adenomyosis were compared with patients with no pathology and leiomyomatous uteri as controls (n = 4 in each group, respectively). Additionally, midsecretory phase endometrial sections were obtained from patients with adenomyosis and control patients with leiomyoma (n = 8 in each group, respectively).
GATA2 and GATA6 immunohistochemistry and H-SCORE were performed on the midsecretory phase endometrial sections from adenomyosis and leiomyoma control patients (n = 8 each, respectively). Control and adenomyosis patient HESC cultures were treated with placebo or 10^-8 M estradiol (E2), or decidualization media (EMC) containing 10^-8 M E2, 10^-7 M medroxyprogesterone acetate, and 5 × 10^-5 M cAMP for 6 and 10 days. Additionally, control HESC cultures (n = 4) were transfected with scrambled small interfering RNA (siRNA) (control) or GATA2-specific siRNAs for 6 days while adenomyosis HESC cultures (n = 4) were transfected with human GATA2 expression vectors to silence or induce GATA2 overexpression.
Immunohistochemistry was performed to obtain GATA2 and GATA6 H-SCORES in adenomyosis vs. control patient endometrial tissue. Expression of GATA2, GATA6, insulin-like growth factor-binding protein 1 (IGFBP1), prolactin (PRL), progesterone receptor (PGR), estrogen receptor 1 (ESR1), leukemia inhibitory factor (LIF), and Interleukin receptor 11 (IL11R) messenger RNA (mRNA) levels were analyzed using by qPCR with normalization to ACTB. Silencing and overexpression experiments also had the corresponding mRNA levels of the above factors analyzed. Western blot analysis was performed on isolated proteins from transfection experiments.
Immunohistochemistry revealed an overall fourfold lower GATA2 and fourfold higher GATA6 H-SCORE level in the endometrial stromal cells of patients with adenomyosis vs. controls. Decidual induction with EMC resulted in significantly lower GATA2, PGR, PRL and IGFBP1 mRNA levels in HESC cultures from patients with adenomyosis patient vs. controls. Leukemia inhibitory factor and IL11R mRNA levels were also significantly dysregulated in adenomyosis HESCs compared with controls. . Silencing of GATA2 expression in control HESCs induced an adenomyosis-like state with significant reductions in GATA2, increases in GATA6 and accompanying aberrations in PGR, PRL, ESR1 and LIF levels. Conversely, GATA2 overexpression via vector in adenomyosis HESCs caused partial restoration of the defective decidual response with significant increases in GATA2, PGR, PRL and LIF expression.
In-vivo and in-vitro experiment results demonstrate that there is an overall inverse relationship between endometrial GATA2 and GATA6 levels in patients with adenomyosis who have diminished GATA2 levels and concurrently elevated GATA6 levels. Additionally, lower GATA2 and higher GATA6 levels, together with aberrant levels of important receptors and implantation factors, such as ESR1, PGR, IGFBP1, PRL, LIF, and IL11R mRNA in HESCs from patients with adenomyosis or GATA2-silenced control HESCs, support impaired decidualization. These effects were partially restored with GATA2 overexpression in adenomyosis HESCs, demonstrating a potential therapeutic target.
Fertility preservation before hematopoetic stem cell transplantation: a case series of women with GATA binding protein 2 deficiency, dedicator of cytokinesis 8 deficiency, and sickle cell disease
2020, F and S Reports
Citation Excerpt :
Our study demonstrates the feasibility of oocyte cryopreservation in patients with GATA2 deficiency and DOCK8 deficiency, two rare single-gene disorders that impact immune function. GATA binding protein 2 expression has been observed in epithelial and stromal tissues of the uterus and is correlated positively with P receptor levels (14, 15). Mutations in GATA2 have been hypothesized to alter P responsiveness through reducing P receptor expression, which may manifest clinically as failed embryo implantation, impaired endometrial decidualization, and uninhibited estrogen signaling (15, 16).
To describe fertility characteristics, outcomes of oocyte cryopreservation cycles, and safety of ovarian stimulation in patients with GATA binding protein 2 (GATA2) deficiency, dedicator of cytokinesis 8 (DOCK8) deficiency, and sickle cell disease (SCD) preparing for hematopoetic stem cell transplantation (HSCT).
Retrospective case series.
The National Institutes of Health.
Female patients with GATA2 deficiency, DOCK8 deficiency, and SCD aged between 13 and 38 years.
None.
Demographic and ovarian reserve parameters, stimulation outcomes, and adverse event occurrences were collected through chart review. Descriptive statistics were used to identify trends within disease subcategories.
Twenty-one women with GATA2 deficiency, DOCK8 deficiency, and SCD underwent fertility preservation prior to HSCT. Patients with DOCK8 deficiency had the lowest mean age (16.5 years old) and antimüllerian hormone (0.85 ng/mL). Patients with GATA2 deficiency had the highest antral follicle count and antimüllerian hormone (25.77 and 5.07 ng/mL, respectively). Baseline follicle-stimulating hormone, luteinizing hormone, and estradiol were comparable between the cohorts. The duration of stimulation was similar (10.43 to 11.25 days) across all groups. Comparable peak estradiol levels were achieved across the cohorts. Patients with SCD had the highest mature (MII) oocyte yield (10.71). Three patients experienced complications related to stimulation: pain crisis in a patient with SCD, pulmonary embolism, and zero oocytes cryopreserved in a patient with GATA2 deficiency.
This study offers insight into controlled ovarian stimulation in patients with these conditions prior to HSCT. Oocyte cryopreservation can be performed successfully, although adverse events must be considered. Following the outcomes of gamete use in this cohort will serve to further our knowledge of the true reproductive potential of this population.
Postnatal exposure to endosulfan affects uterine development and fertility
2020, Molecular and Cellular Endocrinology
Citation Excerpt :
In human endometrial stromal cells, Gata-3 gene expression was demonstrated to be under hormonal conditions mimicking decidualization (Lu et al., 2013). Moreover, Rubel et al. (2012) showed that Gata-2 is expressed during critical phases of early pregnancy, and that it may play a major role in mediating Pg signaling in the mouse uterus. Finally, the E47 TF has been also suggested as a potential regulator of genes associated to endometrial receptivity, by mean of bioinformatic analyses (Tapia et al., 2011).
Endosulfan is an organochlorine pesticide (OCP) used in large-scale agriculture for controlling a variety of insects and mites that attack food and non-food crops. Although endosulfan has been listed in the Stockholm Convention as a persistent organic pollutant to be worldwide banned, it is still in use in some countries. Like other OCPs, endosulfan is bioaccumulative, toxic and persistent in the environment. Human unintentional exposure may occur through air inhalation, dietary, skin contact, as well as, via transplacental route and breast feeding. Due to its lipophilic nature, endosulfan is rapidly absorbed into the gastrointestinal tract and bioaccumulates in the fatty tissues.
Similar to other OCPs, endosulfan has been classified as an endocrine disrupting chemical (EDC). Endocrine action of endosulfan on development and reproductive function of males has been extensively discussed; however, endosulfan effects on the female reproductive tract have received less attention. This review provides an overview of: i) the fate and levels of endosulfan in the environment and human population, ii) the potential estrogenic properties of endosulfan in vitro and in vivo, iii) its effects on uterine development, and iv) the long-term effects on female fertility and uterine functional differentiation during early gestation.
Structure, Function, and Evaluation of the Female Reproductive Tract
2019, Yen & Jaffe's Reproductive Endocrinology: Physiology, Pathophysiology, and Clinical Management: Eighth Edition
The endometrium develops within the uterus, originating from the urogenital ridge early in development. This mucosal lining consists of epithelial and stromal cells invested with a specialized vasculature that matures at puberty and orchestrates cyclic menses while providing an immunoprotected site for implantation of the fetal allograft. As with other hormone responsive proliferative tissues, such as breast or prostate, the endometrium is prone to pathology. In this chapter we review the physiology of this tissue and explore the mechanisms of the normal and abnormal menstrual cycle. The mechanisms of implantation and menstruation are discussed along with the clinical approach to the evaluation of the endometrium in health and disease.
An Ancient Fecundability-Associated Polymorphism Creates a GATA2 Binding Site in a Distal Enhancer of HLA-F
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Similar to the DeepSea results, substituting the reference A allele with the alternative G allele is predicted to abolish GATA2 binding (Figures 2B and 2C). These results suggest that the G/A polymorphism alters binding of GATA2, a transcription factor that plays an essential role in mediating the transcriptional response to progesterone in DSCs.49–51 Previous studies have shown that HLA-F is expressed in placental villi, EVTs, and DSCs.17–19,52
Variation in female reproductive traits, such as fertility, fecundity, and fecundability, are heritable in humans, but identifying and functionally characterizing genetic variants associated with these traits have been challenging. Here, we explore the functional significance and evolutionary history of a G/A polymorphism at SNP rs2523393, which is an eQTL for HLA-F and is significantly associated with fecundability (the probability of being pregnant within a single menstrual cycle). We replicated the association between the rs2523393 genotype and HLA-F expression by using GTEx data and demonstrate that HLA-F is upregulated in the endometrium during the window of implantation and by progesterone in decidual stromal cells. Next, we show that the rs2523393 A allele creates a GATA2 binding site in a progesterone-responsive distal enhancer that loops to the HLA-F promoter. Remarkably, we found that the A allele is derived in the human lineage and that the G/A polymorphism arose before the divergence of modern and archaic humans and segregates at intermediate to high frequencies across human populations. Remarkably, the derived A allele is has also been identified in a GWAS as a risk allele for multiple sclerosis. These data suggest that the polymorphism is maintained by antagonistic pleiotropy and a reproduction-health tradeoff in human evolution.
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Progesterone acts through the progesterone receptor to direct physiological adaption of the uterus in preparation and completion of pregnancy. Genome-wide transcriptome and cistrome analyses have uncovered new members and novel modifiers of the progesterone signaling pathway. Genetically engineered mice allow functional assessment of newly identified genes in vivo and provide insights on the impact of progesterone receptor-dependent molecular mechanisms on pregnancy at the organ system level. Progesterone receptor isoforms collectively mediate progesterone signaling via their distinct and common downstream target genes, which makes the stoichiometry of isoforms relevant in modifying the progesterone activity. This review discusses recent advances on the discovery of the progesterone receptor network, with special focus on the endometrium at early pregnancy and myometrium during parturition.

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GATA2 is expressed at critical times in the mouse uterus during pregnancy

Abstract

Highlights

Section snippets

Confirmation of Gata2 as a progesterone target gene in the murine uterus

Transfection and luciferase assay

Acknowledgements

Fertil Steril

Semin. Cell Dev. Biol.

J. Biol. Chem.

Dev. Biol.

Dev. Biol.

J. Biol. Chem.

Mol. Cell Endocrinol.

Progesterone: how did the name originate?

South Med. J.

Determination of the transcript profile of human endometrium

Mol. Hum. Reprod.

Temporal expression profiling of the uterine luminal epithelium of the pseudo-pregnant mouse suggests receptivity to the fertilized egg is associated with complex transcriptional changes

Hum. Reprod.

Identification of murine uterine genes regulated in a ligand-dependent manner by the progesterone receptor

Endocrinology

Global gene profiling in human endometrium during the window of implantation

Endocrinology

DNA-binding specificities of the GATA transcription factor family

Mol. Cell Biol.

Indian hedgehog is a major mediator of progesterone signaling in the mouse uterus

Nat. Genet.

Bmp2 is critical for the murine uterine decidual response

Mol. Cell Biol.

Molecular evolution of the GATA family of transcription factors: conservation within the DNA-binding domain

J. Mol. Evol.

Mice lacking progesterone receptor exhibit pleiotropic reproductive abnormalities

Genes Dev.