ToxSci Advance Access originally published online on January 5, 2005
Toxicological Sciences 2005 84(2):270-277; doi:10.1093/toxsci/kfi072
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Estrogen Receptor 
Pathway Is Involved in the Regulation of Calbindin-D9k in the Uterus of Immature Rats
* Laboratory of Veterinary Biochemistry and Molecular Biology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, 361-763 Republic of Korea, and Department of Obstetrics and Gynecology, British Columbia Children's and Women's Hospital, British Columbia Research Institute for Children's and Women's Health, University of British Columbia, Vancouver, BC, V6H 3V5 Canada
Received November 8, 2004; accepted December 22, 2004
It has been demonstrated in our previous studies that Calbindin-D9k (CaBP-9k) is a potent biomarker for screening estrogen-like chemicals in the rat model. Although treatments with 17beta-estradiol (E2) and endocrine disrupting compounds resulted in the up-regulation of uterine CaBP-9k, the mechanism of CaBP-9k induction by these compounds through two subtypes of estrogen receptors (ER
and ERß) is unclear. Thus, in the present study, immature rats were treated with propyl pyrazole triol (PPT, an ER-selective ligand), diarylpropionitrile (DPN, an ERß-selective ligand), E2, or dimethyl sulfoxide (DMSO, a vehicle control) for three days in order to clarify which subtype of ER is involved in the uterine CaBP-9k induction. Following injection with these ER ligands, uterine CaBP-9k expression was analyzed by Northern blot and immunoblot assays. Uterine CaBP-9k expression is mainly mediated by PPT in a dose- and time-dependent manner in immature rats, whereas no significant alteration of the uterine CaBP-9k gene was observed after DPN treatment. In addition, an estrogenicity of PPT in inducing CaBP-9k expression was completely blocked by the anti-estrogen ICI 182,780, implying that uterine CaBP-9k is solely induced by ER. A single treatment with PPT rapidly increased the protein levels of ER and PR, an E2-mediated gene, in these tissues. Taken together, these results indicate that uterine CaBP-9k is induced by E2 and endocrine disrupting chemicals via the ER pathway, but not ERß, in the uterus of immature rats.
Key Words: Calbindin-D9k; estrogen receptor; progesterone receptor; PPT; DPN.
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