Unknown,Transcriptomics,Genomics,Proteomics

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Effects of estrogen and progesterone on glycosyltransferase expression in mice

ABSTRACT: This laboratory studies the structural basis of carbohydrate function in terms of adhesion and other biological processes. The Fisher lab is testing the hypothesis that the mucin-coated oral and uterine cavities present similar carbohydrate receptors that specify the bacterial ecology of both regions and the repertoire of these oligosaccharide species is hormonally regulated. This theory also suggests that certain individuals express carbohydrate receptors that make them susceptible to both periodontal disease and preterm labor. Experimental procedure: 40 mature female mice were ovariectomize . The mice were allowed to rest for two weeks to eliminate any remaining endogenous estrogen (E2) and progesterone (P4). Then the mice were separated into four groups (10 each) that received the treatment indicated. Group 1: ovariectomized mice with no supplement, only sesame oil vehicle. Group 2: ovariectomized mice with only P4 supplement (2 mg/day/mouse). Group 3: ovariectomized mice with E2 supplement (100 ng/day/mouse). Group 4: ovariectomized mice with both P4 (2 mg/day/mouse) and E2 (100 ng/day/mouse) supplement. The treatment lasted for 4 days; the steroids were dissolved in sesame oil and injected subcutaneously. After treatment, salivary glands (parotid, submandibular, and sublingual gland), submandibular lymph nodes and uterine horns with cervix were collected. Total RNA was extracted from these samples using Trizol.

ORGANISM(S): Mus musculus

SUBMITTER: Steven Head

PROVIDER: E-GEOD-27598 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Ovarian estrogen (E2) and progesterone (P4) are indispensable for embryo-implantation and endometrial stromal decidualization; however, the molecular mechanisms that underpin these reproductive processes are unclear. Steroid receptor coregulator-2 (SRC-2) belongs to the multifunctional SRC/p160 family which also includes SRC-1 and SRC-3. Sharing strong sequence homology, all three SRCs exert diverse regulatory effects by modulating the transcriptional potency of nuclear receptor family members, including the estrogen and progesterone receptor (ER and PR respectively). Importantly, absence of SRC-2 in PR positive cells in the epithelial, stromal, and myometrial compartments of the murine uterus results in a striking infertility defect. This reproductive phenotype highlights a key role for SRC-2 in uterine function which is not shared with other coregulators. Intriguingly, abrogation of uterine SRC-2 does not block embryo apposition or attachment to the apical surface of luminal epithelial cells of the endometrium but rather prevents P4-dependent local decidualization of the sub-epithelial stroma. Remarkably, epithelial-specific ablation of SRC-2 in the murine uterus does not compromise endometrial functionality, again underscoring the unique importance of stromal derived SRC-2 in uterine function. The stromal decidualization defect resulting from SRC-2 ablation is reflected at the molecular level by a marked attenuation in P4 responsive target genes known to be critical for P4 dependent decidualization (i.e. ERBB receptor feedback inhibitor 1, Follistatin and Fkbp5). Conversely, the induction of E2 or P4 target genes involved in embryo implantation (i.e. leukemia inhibitory factor (LIF) and Indian hedgehog (Ihh) respectively) is not affected by SRC-2’s absence. As with mouse studies, decidualization of primary human stromal cells (HESCs) in culture is blocked by SRC-2 knockdown; however, HESC decidualization is unaffected by knockdown of SRC-1 or SRC-3. As a consequence of SRC-2 knockdown, molecular studies disclose a striking decrease in the induction of a subset of P4 target genes (i.e. WNT4 and FKBP5) which are essential for the stromal-epithelioid transformation step, the cellular hallmark of endometrial decidualization. Collectively, these studies not only showcase the evolutionary importance of SRC-2 in endometrial biology but also suggest that deregulation of this coregulator may underpin a spectrum of hormone-dependent uterine pathologies such as endometriosis and endometrial cancer. Microarray analysis was performed on mouse uteri using eighteen SRC-2flox/flox (SRC-2f/f) and eighteen PRCre/+ SRC-2flox/flox (SRC-2d/d) mice. Mice were ovariectomized at 6 weeks and after 2 weeks mice were either treated with sesame oil (vehicle) or 1 mg of P4. RNA from three mice per genotype per treatment were pooled and assigned as one sample (three samples per genotype per treatment). multiple group comparison

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Effects of estrogen and progesterone on glycosyltransferase expression in mice

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