191 ESTROGEN REGULATES THE LOCALIZATION AND EXPRESSION OF CALBINDIN-D9k IN THE PITUITARY GLAND OF IMMATURE MALE RATS VIA THE ESTROGEN RECEPTOR ALPHA PATHWAY

Abstract

Estrogen (E2; estradiol) plays a key role in the regulation of many pituitary hormones. It exerts its effects by binding to the intracellular estrogen receptor (ER), which then functions as a transcription factor. Although E2 has been shown to regulate calbindin-D9k (CaBP-9k) in the female reproductive system of rodents, the effects of E2 on the regulation of CaBP-9k in male rats remains to be elucidated. For investigation of E2-induced regulation of the pituitary CaBP-9k gene, immature male rats were injected with E2 daily for three consecutive days with a dose of 40 µg kg^–1 body weight (BW). The expression levels of CaBP-9k mRNA and protein were analyzed by RT-PCR and Western blot analysis, respectively, in the absence and presence of ICI 182 780 (ICI), an E2 antagonist. In addition, the tissue localization of CaBP-9k was determined by immunohistochemistry. CaBP-9k was localized in the cytoplasm of a specific cell type (acidophils) in the anterior lobe of the pituitary gland and highly expressed in the intermediate lobe. Exposure to E2 increased the number of cells that stained positive for CaBP-9k. For determination of which ER subtype is involved in CaBP-9k regulation in the pituitary, the immature rats were treated with 1 mg kg^–1 BW propyl pyrazole triol (PPT, an ER-α-selective ligand) or diarylpropionitrile (DPN, an ER-β-selective ligand) for three days. The data were analyzed by the non-parametric procedure of the Kruskal-Wallis test, followed by Dunnett's test for two-pair comparisons. Pituitary CaBP-9k expression was mainly mediated by PPT in immature male rats, whereas no significant alteration of pituitary CaBP-9k gene expression was observed after DPN treatment. In addition, the estrogenicity of PPT in the induction of CaBP-9k expression was completely blocked by an estrogen antagonist, ICI 182 780, indicating that pituitary CaBP-9k expression is solely induced by ER-α. Taken together, these results suggest that pituitary CaBP-9k is induced by estrogen in male rats and its expression is predominantly regulated by ER-α, but not ER-β.