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Ventromedial Hypothalamic Nucleus-Specific Enhancer of Ad4BP/SF-1 Gene

Division for Sex Differentiation (Y.S., M.Z., S.I., Y.S., S.Oka, K-i.M.), National Institute for Basic Biology, National Institutes of Natural Sciences, Myodaiji-cho, Okazaki 444-8787, Japan; Core Research for Evolutional Science and Technology (CREST) (Y.S., M.Z., S.I., Y.S., S.Oka, K.-i.M.), Japan Science and Technology Corporation, Kawaguchi 332-0012, Japan; Department of Pediatric Surgery (Y.S., S.S.), Reproductive and Developmental Medicine, Graduate School of Medical Sciences, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan; Laboratory of Metabolism (S.K.), National Institutes of Health, Bethesda, Maryland 20892; and Division of Endocrinology and Metabolism (S.Okam., Y.M.), Department of Developmental Physiology, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Myodaiji-cho, Okazaki 444-8585, Japan

Address all correspondence and requests for reprints to: Prof. Ken-ichirou Morohashi, PhD., Division for Sex Differentiation, National Institute for Basic Biology, National Institutes of Natural Sciences, Myodaiji-cho, Okazaki 444-8787, Japan. E-mail: moro{at}nibb.ac.jp.

Ad4BP/SF-1 [Ad4 binding protein/steroidogenic factor-1 (designated NR5A1)] is a transcription factor essential for animal reproduction. Based on the phenotypes observed in gene-disrupted mice, Ad4BP/SF-1 is thought to be involved in establishment of the hypothalamic-pituitary-gonadal axis. However, the mechanisms underlying tissue-specific expression of Ad4BP/SF-1 are largely unknown. Here, we investigated the cis-regulatory regions of the mouse Ad4BP/SF-1 gene by transgenic mouse assays, and identified a ventromedial hypothalamic nucleus (VMH)-specific enhancer. The enhancer localized in intron 6 is highly conserved between mouse, human, and chick. The enhancer has the potential to reproduce endogenous gene expression from the fetal ventromedial diencephalon to the adult VMH. The VMH enhancer was characterized by the presence of suppressive and activating elements. Mutation of the former element resulted in ectopic lacZ reporter gene expression in an area dorsal to the intrinsic expression domain and in the ventricular zone, whereas mutations in the latter containing ATTA motifs led to the disappearance of the reporter gene expression, suggesting the involvement of homeobox proteins. Using nuclear extracts prepared from the adult hypothalami, EMSAs identified specific protein binding to the activating elements but not to the suppressive element.

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Nuclear Receptors:   SF-1

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