Characterization of transgenic mice expressing EGFP under the control of the monkey 20α-hydroxysteroid dehydrogenase promoter
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To directly assess the molecular function of the monkey 20α-hydroxysteroid dehydrogenase (20α-HSD) promoter, we generated transgenic mice (tg) expressing enhanced green fluorescent protein (EGFP) under control of this promoter. We demonstrated that prostaglandin F2α induced 20α-HSD promoter activity in CHO cells in a dose-dependent manner. Furthermore, forskolin treatment markedly reduced 20α-HSD promoter activity, and prolactin exhibited weak inhibitory activity. The transgenic mouse obtained one positive founder male. The transgene was propagated in 10 successive generations without any notable defects to the progeny. EGFP and 20α-HSD in the tg mice were colocalized in the luteal cells of the ovary during late pregnancy. Strong EGFP and 20α-HSD protein signals were also detected in the adult testis. Immunohistochemical analysis revealed high EGFP levels in the seminiferous epithelium, whereas 20α-HSD was expressed in the seminiferous tubules. Our data suggest that the ovaries in monkey and mouse exhibit similar expression patterns of 20α-HSD during pregnancy. However, the expression pattern of EGFP in tg mice testis slightly differed from that of the endogenous 20α-HSD. Further investigation is required to elucidate the functional mechanisms underlying regulation of the monkey 20α-HSD promoter in the tg mice.
Keywordstransgenic mice EGFP monkey 20α-HSD promoter
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The authors thank Dr. HW Seong (Institute of Animal Science) for his helpful discussions. This study was supported by the KRIBB Research Initiative Program (KGM4251622), Republic of Korea.
- Albarracin, C.T., Parmer, T.G., Duan, W.R., Nelson, S.E., and Gibori, G. (1994). Identification of a major prolactin-regulated protein as 20 alphahydroxysteroid dehydrogenase: coordinate regulation of its activity, protein content, and messenger ribonucleic acid expression. Endocrinology 134, 2453–2460.CrossRefPubMedGoogle Scholar
- Madore, E., Harvey, N., Parent, J., Chapdelaine, P., Arosh, J.A., and Fortier, M.A. (2003). An aldose reductase with 20α-hydroxysteroid dehydrogenase activity is most likely the enzyme responsible for the production of prostaglandin F2α in the bovine endometrium. J Biol Chem 278, 11205–11212.CrossRefPubMedGoogle Scholar
- Naidansuren, P., Park, C.W., Kim, S.H., Nanjidsuren, T., Park, J.J., Yun, S. J., Sim, B.W., Hwang, S., Kang, M.H., Ryu, B.Y., et al. (2011). Molecular characterization of bovine placental and ovarian 20α-hydroxysteroid dehydrogenase. Reproduction 142, 723–731.CrossRefPubMedPubMedCentralGoogle Scholar
- Nanjidsuren, T., Naidansuren, P., Park, C.W., Park, J.J., Yun, S.J., Sim, B. W., Kang, M.H., Lee, S.R., Chang, K.T., and Min, K.S. (2011). Expression and localization of the 20α-hydroxysteroid dehydrogenase (HSD) enzyme in the reproductive tissues of the cynomolgus monkey Macaca fascicularis. J Steroid Biochem Mol Biol 127, 337–344.CrossRefPubMedGoogle Scholar
- Seo, K.S., Naidansuren, P., Kim, S.H., Yun, S.J., Park, J.J., Sim, B.W., Park, C.W., Nanjidsuren, T., Kang, M.H., Seo, H., et al. (2011). Expression of aldo-keto reductase family 1 member C1 (AKR1C1) gene in porcine ovary and uterine endometrium during the estrous cycle and pregnancy. Reprod Biol Endocrinol 9, 139.CrossRefPubMedPubMedCentralGoogle Scholar
- Sreenivasulu, G., Senthilkumaran, B., Sridevi, P., Rajakumar, A., and Rasheeda, M.K. (2012). Expression and immunolocalization of 20β-hydroxysteroid dehydrogenase during testicular cycle and after hCG induction, in vivo in the catfish, Clarias gariepinus. General Comp Endocrinol 175, 48–54.CrossRefGoogle Scholar
- Sudeshna, T., Anand, K., and Medhamurthy, R. (2013). Analysis of 20alpha-hydroxysteroid dehydrogenase expression in the corpus luteum of the buffalo cow: effect of prostaglandin F2-alpha treatment on circulating 20alpha-hydroxyprogesterone levels. Reprod Biol Endocrinol 11, 111.CrossRefPubMedPubMedCentralGoogle Scholar