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Abstract

Volume 9, Issue 4 (July 2007) 9, 515–521; 10.1111/j.1745-7262.2007.00300.x

Epididymis-specific lipocalin promoters

Kichiya Suzuki, Xiuping Yu, Pierre Chaurand, Yoshihiko Araki, Jean-Jacques Lareyre, Richard M Caprioli, Marie-Claire Orgebin-Crist and Robert J Matusik

1.Department of Obstetrics and Gynecology, Tohoku University Graduate School of Medicine, Sendai 980-8574, Japan
2.Department of Urologic Surgery, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA
3.Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA
4.Department of Obstetrics and Gynecology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA
5.Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA
6.Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA
7.Center for Reproductive Biology Research, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA

Correspondence: Dr Kichiya Suzuki, Department of Obstetrics and Gynecology, Tohoku University Graduate School of Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai 980-8574, Japan. Fax: +81-22-717-7258. E-mail: kichiyasuzuki@mail.tains.tohoku.ac.jp

Abstract

Our goal is to decipher which DNA sequences are required for tissue-specific expression of epididymal genes. At least 6 epididymis-specific lipocalin genes are known. These are differently regulated and regionalized in the epididymis. Lipocalin 5 (Lcn5 or mE-RABP) and Lipocalin 8 (Lcn8 or mEP17) are homologous genes belonging to the epididymis-specific lipocalin gene cluster. Both the 5 kb promoter fragment of the Lcn5 gene and the 5.3 kb promoter fragment of the Lcn8 gene can direct transgene expression in the epididymis (Lcn5 to the distal caput and Lcn8 to the initial segment), indicating that these promoter fragments contain important cis-regulatory element(s) for epididymis-specific gene expression. To define further the fragments regulating gene expression, the Lcn5 promoter was examined in transgenic mice and immortalized epididymal cell lines. After serial deletion, the 1.8 kb promoter fragment of the Lcn5 gene was sufficient for tissue-specific and region-specific gene expression in transgenic mice. Transient transfection analysis revealed that a transcription factor forkhead box A2 (Foxa2) interacts with androgen receptor and binds to the 100 bp fragment of the Lcn5 promoter between 1.2 kb and 1.3 kb and that Foxa2 expression inhibits androgen-dependent induction of the Lcn5 promoter activity. Immunohistochemistry indicated a restricted expression of Foxa2 in the epididymis where endogenous Lcn5 gene expression is suppressed and that the Foxa2 inhibition of the Lcn5 promoter is consistent with the lack of expression of Lcn5 in the corpus and cauda. Our approach provides a basic strategy for further analysis of the epididymal lipocalin gene regulation and flexible control of epididymal function.

Keywords: androgen, epididymis, gene cluster, gene evolution, gene regulation, lipocalin, transcription factor

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