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Zebrafish Androgen Receptor: Isolation, Molecular, and Biochemical Characterization¹

Reproductive Genomics Group,⁵ Temasek Life Sciences Laboratory, Singapore 117604, Singapore Department of Biological Sciences,⁶ National University of Singapore, Singapore 117543, Singapore Unit of Molecular Biology,⁷ Department of Natural Sciences, Örebro University, SE-701 82 Örebro, Sweden

ABSTRACT

Androgens play an important role in male sexual differentiation and development. They exert their function by binding to and activating the androgen receptor (Ar), a member of the steroid hormone receptor superfamily. Here, we report on the isolation and characterization of zebrafish Ar. The complete transcript of zebrafish ar is 5.3 kb long encoding a putative polypeptide of 868 amino acids. Our experimental and bioinformatic analysis has found a single ar locus in zebrafish. Phylogenetic analysis using the ligand-binding domain showed that the zebrafish Ar clustered with its cyprinid orthologs to form a separate group, which was closer to the beta clade than to the alpha clade. Tissue-specific expression analysis revealed that the ar mRNA was expressed ubiquitously in all adult tissues tested, with sexually dimorphic expression in the gonad and muscle. While the ar transcript was maternally deposited into the embryo, signs of zygotic expression could be detected as early as 24 h after fertilization, and the expression level increased substantially afterwards. When analyzed during gonad development, the expression level of ar mRNA at 4 wk after fertilization was similar in both developing gonads but later became higher in the transforming testis, suggesting a potential role during male gonad differentiation. We also combined theoretical modeling with in vitro experiments to show that the zebrafish Ar is preferentially activated by 11-ketotestosterone.

11-ketotestosterone, androgen receptor, gonad differentiation, steroid hormones, testis

⁴These authors contributed equally to this work.

¹Supported by grants from the Swedish Animal Welfare Agency (PEO) and the Swedish Knowledge Foundation (PEO), and by an internal research grant from Temasek Life Sciences Laboratory, Singapore.

Correspondence: ²Laszlo Orban, Reproductive Genomics Group, Temasek Life Sciences Laboratory, 1 Research Link, The NUS, Singapore 117604. FAX: 65 6872 7007; e-mail: laszlo{at}tll.org.sg

Correspondence: ³Per-Erik Olsson, Örebro Life Science Center, Department of Natural Sciences, Örebro University, SE-701 82 Örebro, Sweden. FAX: 46 19 303566; e-mail: per-erik.olsson{at}nat.oru.se

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				P P de Waal, D S Wang, W A Nijenhuis, R W Schulz, and J Bogerd Functional characterization and expression analysis of the androgen receptor in zebrafish (Danio rerio) testis Reproduction, August 1, 2008; 136(2): 225 - 234. [Abstract] [Full Text] [PDF]