Molecular cloning of DAX1 and SHP cDNAs and their expression patterns in the Nile tilapia, Oreochromis niloticus

Wang, De Shou; Kobayashi, Tohru; Senthilkumaran, B.; Sakai, Fumie; Sudhakumari, C.C.; Suzuki, Tsuneyuki; Yoshikuni, Michiyasu; Matsuda, Masaru; Morohashi, Ken Ichirou; Nagahama, Yoshitaka

doi:10.1016/s0006-291x(02)02252-0

Cited by 64 publications

(43 citation statements)

References 21 publications

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“…The RT-PCR cycles were as follows: 94 C for 2 min, followed by 30 to 36 cycles of 94 C (30 s), 60 C (30 s), and 72 C (1 min), ending with 10 min of extension at 72 C. Positive and negative controls were set up with respective plasmid DNA and water as templates to validate the distribution patterns. A 342 bp tilapia -actin fragment was amplified (as internal control) to test the quality of the cDNA used in the PCR reactions (Wang et al 2002). All the PCR products were electrophoresed using 1·5% agarose gels and the gels were stained with ethidium bromide to visualize bands.…”

Section: Tissue Distribution Analysis Of Three Types Of 17 −Hsds By Rmentioning

confidence: 99%

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Cloning, expression and characterization of three types of 17β-hydroxysteroid dehydrogenases from the Nile tilapia, Oreochromis niloticus

Zhou¹,

Wang²,

Senthilkumaran³

et al. 2005

Journal of Molecular Endocrinology

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In order to elucidate the roles of 17 -HSDs in fish gonadal steroidogenesis, three types of 17 -HSDs (17 -HSD1, 17 -HSD8 and putative 17 -HSD12) were cloned and characterized from the Nile tilapia, Oreochromis niloticus. The cloned cDNAs of 17 -HSD type 1, 8 and 12 were 1504, 1006 and 1930 bp long, with open reading frames encoding proteins of 289, 256 and 314 aminoacids, respectively. Tissue distribution pattern analyzed by RT-PCR and Northern blot showed that 17 -HSD1 was dominantly expressed in the ovary, while the putative 17 -HSD12, one of the two duplicates found in fish, is a male specific enzyme and expressed exclusively in testis (detected by RT-PCR only). On the other hand, 17 -HSD8 was expressed in the brain, gill, heart, liver, intestine, gonad, kidney and muscle of both male and female. Enzymatic assays of the three types of 17 -HSDs were performed using recombinant proteins expressed in E. coli or HEK 293 cells. Tilapia 17 -HSD1 expressed in E. coli had the preference for NADP(H) as cofactor and could catalyze the inter-conversion between estrone and estradiol efficiently as well as the inter-conversion between androstenedione and testosterone, but less efficiently. Tilapia 17 -HSD8 recombinant protein expressed in HEK 293 cells could catalyze the conversion of testosterone to androstenedione, as well as the inter-conversion between estrone and estradiol. However, the putative 17 -HSD12 expressed in E. coli or in HEK 293 cells showed no conversion to any of the four substrates tested in this study. Based on enzyme characterization and tissue distribution, it is plausible to attribute crucial roles to 17 -HSDs in the gonadal steroidogenesis of teleosts.

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Section: Tissue Distribution Analysis Of Three Types Of 17 −Hsds By Rmentioning

confidence: 99%

“…Northern blot was performed as per the method reported previously (Wang et al 2002, Jiang et al 2003. Briefly, total RNA was extracted from various tissues, such as gonad, liver, intestine and brain of both male and female fish using ISOGEN solution as per the manufacturer's protocol (Nippon Gene, Toyama, Japan).…”

Section: Northern Blot Analysismentioning

confidence: 99%

Cloning, expression and characterization of three types of 17β-hydroxysteroid dehydrogenases from the Nile tilapia, Oreochromis niloticus

Zhou¹,

Wang²,

Senthilkumaran³

et al. 2005

Journal of Molecular Endocrinology

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“…Several transcription factors like sry, dmy, sox9, ad4BP/SF-1, dmrt1, dax1 and wt1 have been identified to play an important role in sex determination and differentiation of vertebrates including teleosts (Swain & Lovell-Badge 1999, Hughes 2001, Wang et al 2002, Kobayashi et al 2004. Molecular similarity in sexual development across different phyla found so far is among Drosophila doublesex, Caenorhabditis mab-3 and vertebrate dmrt1 (Raymond et al 1998).…”

Section: Introductionmentioning

confidence: 99%

Identification of multiple dmrt1s in catfish: localization, dimorphic expression pattern, changes during testicular cycle and after methyltestosterone treatment

Kavarthapu

Senthilkumaran²

2009

Journal of Molecular Endocrinology

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The double sex and mab-3 related (DM) transcription factor 1 (dmrt1) plays an important role in testicular differentiation. Here, we report cloning of multiple dmrt1s, a full-length and two alternative spliced forms from adult catfish (Clarias gariepinus) testis, which encode predicted proteins of 287 (dmrt1a), 253 (dmrt1b) and 233 (dmrt1c) amino acid residues respectively. Interestingly, dmrt1c lacks the majority of the DM domain. Multiple dmrt1s (dmrt1a and dmrt1c) were obtained from Clarias batrachus as well. Tissue distribution (transcript and protein) of catfish dmrt1 revealed exclusive expression in testis. Semi-quantitative RT-PCR revealed the presence of multiple dmrt1s with high levels of dmrt1a in adult testis but not in ovary. Real-time RT-PCR analysis during testicular cycle showed higher levels of dmrt1 transcripts in preparatory and pre-spawning when compared with spawning and post-spawning phases. Immunocytochemical and immunofluorescence localization revealed the presence of catfish Dmrt1 protein in spermatogonia and spermatocytes, which indicates plausible role in spermatogenesis. Histological analysis indicated initiation of gonadal sex differentiation in catfish around 40-50 days after hatching. The potential role for dmrt1 in testicular differentiation is evident from its stage-dependent elevated expression in developing testis. Furthermore, dimorphic expressions of dmrt1s were evident at different stages of gonadal development or recrudescence in catfish. Treatment of methyl testosterone (MT) during early stages of gonadal sex differentiation resulted in adult males. Interestingly, we also obtained MT-treated fishes having ova-testis gonads. Analysis of dmrt1, sox9a, foxl2 and cyp19a1 expression patterns in MT-treated gonads revealed tissue-specific pattern. These results together suggest that multiple dmrt1s are testis-specific markers in catfish.

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“…Inside each repeat a LXXLL motif is present, which is thought to have an important role in the mechanism of transcriptional regulation by DAX-1 (see Section 4). DAX-1 homologues have been cloned in a variety of species [6][7][8][9][10][11][12]. Remarkably, nonmammalian DAX-1 homologues have a much shorter N-terminal domain, harboring only one repeat.…”

Section: Cloning and Structure Of Dax-1: An Unusual Orphan Receptormentioning

confidence: 99%

“…Furthermore, in humans DAX-1 is expressed both in the male and the female embryonic gonads during the critical period of sex determination [22]. It is also worth to be mentioned that DAX-1 homologues are expressed during embryogenesis in the gonads of both sexes in another mammalian species (pig) and in chicken, frog, and fish [7,8,10,11]. Moreover, in alligator and turtle species where sexual differentiation is determined by temperature, Dax-1 is expressed at similar levels in male and female gonads during the critical period for their specification [9,12].…”

Section: Dax-1 Role In Sex Determination and Gonadal Function: Lessonmentioning

confidence: 99%

Role of Orphan Nuclear Receptor DAX-1/NR0B1 in Development, Physiology, and Disease

Lalli

2014

Advances in Biology

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is an unusual orphan receptor that has a pivotal role in the development and function of steroidogenic tissues and of the reproductive axis. Recent studies have also indicated that this transcription factor has an important function in stem cell biology and in several types of cancer. Here I critically review the most important findings on the role of DAX-1 in development, physiology, and disease of endocrine tissues since the cloning of its gene twenty years ago.

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Molecular cloning of DAX1 and SHP cDNAs and their expression patterns in the Nile tilapia, Oreochromis niloticus

Cited by 64 publications

References 21 publications

Cloning, expression and characterization of three types of 17β-hydroxysteroid dehydrogenases from the Nile tilapia, Oreochromis niloticus

Cloning, expression and characterization of three types of 17β-hydroxysteroid dehydrogenases from the Nile tilapia, Oreochromis niloticus

Identification of multiple dmrt1s in catfish: localization, dimorphic expression pattern, changes during testicular cycle and after methyltestosterone treatment

Role of Orphan Nuclear Receptor DAX-1/NR0B1 in Development, Physiology, and Disease

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