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Gonadotropin-releasing hormone and gonadal steroids regulate transcription factor mRNA expression in primary pituitary and immortalized gonadotrope cells.

Reproductive sciences (Thousand Oaks, Calif.) (2015-01-08)
Weiming Zheng, Constance M Grafer, Jonathan Kim, Lisa M Halvorson
ABSTRACT

Hormonal regulation of pituitary gonadotropin gene expression has been attributed to gonadotropin-releasing hormone (GnRH)-mediated stimulation of immediate early gene expression and gonadal steroid interactions with their respective nuclear receptors. A number of orphan nuclear receptors including steroidogenic factor 1, liver receptor homologue 1, dosage-sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1, and chicken ovalbumin upstream promoter-transcription factors I/II as well as the GATA family members, GATA2 and GATA4, have also been implicated in transcriptional regulation of the gonadotropin genes. We hypothesized that hormonally mediated changes in these latter transcription factors may provide an additional mechanism for mediating hormonal effects beyond the more classically appreciated pathways. In these studies, we demonstrate significant regulation of orphan nuclear receptor and GATA messenger RNA levels by GnRH, dihydrotestosterone, estradiol, and progesterone in both cultured primary pituitary cells and gonadotrope-derived cell line, LβT2. These results advance our understanding of the complex mechanisms by which GnRH and steroid hormones achieve precise regulation of anterior pituitary function.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Progesterone, ≥99%
Sigma-Aldrich
Progesterone, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Estradiol, meets USP testing specifications
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5α-Androstan-17β-ol-3-one, ≥97.5%
Supelco
Estradiol, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Progesterone, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Progesterone, γ-irradiated, BioXtra, suitable for cell culture
Sigma-Aldrich
Progesterone, meets USP testing specifications
Sigma-Aldrich
Luteinizing hormone releasing hormone, ≥97% (HPLC)
USP
Estradiol, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Luteinizing hormone releasing hormone salmon, ≥97% (HPLC)
USP
Progesterone, United States Pharmacopeia (USP) Reference Standard
Supelco
Progesterone, VETRANAL®, analytical standard
Progesterone for system suitability, European Pharmacopoeia (EP) Reference Standard
Progesterone for peak identification, European Pharmacopoeia (EP) Reference Standard
Progesterone, European Pharmacopoeia (EP) Reference Standard