Cloning and expression of candidate sexual development genes in the cane toad (<i>Bufo marinus</i>)

Abramyan, John; Feng, Chun-Wei; Koopman, Peter

doi:10.1002/dvdy.22055

Cited by 26 publications

(38 citation statements)

References 86 publications

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“…We next performed qRT-PCR analysis of Sf1 and Dax1 expression in ovaries and testes of B. marinus at four developmental time points: metamorph, 20, 30, and 60 mm, as described by Abramyan et al (2009). As we cannot reliably determine the sex of the specimens at the metamorph stage, cDNA was generated from pools of mixed-sex samples.…”

Section: Sex-specific Expression Of Sf1 and Dax1mentioning

confidence: 99%

“…Developmental time points were chosen based on Abramyan et al (2009), where metamorphs represent a presexual differentiation time point, 20 mm represents the time point of sexual differentiation, whereas 30 and 60 mm specimens represent postsexual differentiation time points. All animals were collected and measured snout to urostyle as described by Reading ('91).…”

Section: Animal and Tissue Collectionmentioning

confidence: 99%

“…Each sample was run in triplicate and a DCT calculation was performed according to Bookout and Mangelsdorf (2003). Ribosomal 18S RNA (Rn18S) was used as a calibrator (Abramyan et al, 2009). PCR products were subsequently cloned and sequenced as above to check for specificity and authenticity.…”

Section: Rna Extractions and Cdna Synthesismentioning

confidence: 99%

“…After reverse transcription, qRT-PCR was performed with SYBR Green master mix (Applied Biosystems, Foster City, CA). Primer sets described by Abramyan et al (2009) were used for Sox9, Dmrt1, and p450arom analysis. For bmSf1 and bmDax1, specific primer sets were designed, spanned putative introns larger than 2.6 kb when compared with mouse and chicken (G. gallus): RT-Sf1F-ACACGAGAAACTCAGCATG-3 0 and RT-Sf1R 5 0 -CTGCAGTAAA ATCGTCTGG-3 0 ; RT-Dax1F 5 0 -CAGCAACAAAGAGTATGCCTAT CT-3 0 and RT-Dax1R 5 0 -GCTTCCTGTTGTAATCCTTGGATA-3 0 .…”

Section: Rna Extractions and Cdna Synthesismentioning

confidence: 99%

“…In this study, we investigated the possible physiological role of the BO at the molecular level by cloning and sequencing the B. marinus orthologs of Sf1 and Dax1, and analyzing their expression patterns in parallel to previously identified Sox9, Dmrt1, and p450arom (Abramyan et al, 2009). In addition, we investigated the influence of the adjacent gonads on the expression of p450arom in the BO using ex vivo culture experiments.…”

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confidence: 99%

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Molecular characterization of the Bidder's organ in the cane toad (Bufo marinus)

2010

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In toads, both males and females develop a unique gonadal structure called the Bidder's organ (BO), which resembles ovarian tissue and is attached to the anterior part of the gonad. It is not clear whether the BO is a vestigial organ, or has an endocrine function. In this study, we investigated the expression of the gonadal development genes Dmrt1, Sox9, Sf1, Dax1, and p450arom in the developing BO as compared with the gonads of male and female cane toads. We demonstrate that Sf1, Dax1, and p450arom, key genes involved in vertebrate steroidogenesis, are transcriptionally active in the BO during developmental milestones associated with sexual development and maturation. Furthermore, the pattern of transcriptional activity in the BO is completely independent of the corresponding gonads in both sexes, despite its ovary-like morphology. These results suggest that the BO likely has a steroidogenic role in the development of the cane toad, distinct from that of the gonads.

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Section: Sex-specific Expression Of Sf1 and Dax1mentioning

confidence: 99%

Section: Animal and Tissue Collectionmentioning

confidence: 99%

Section: Rna Extractions and Cdna Synthesismentioning

confidence: 99%

Section: Rna Extractions and Cdna Synthesismentioning

confidence: 99%

mentioning

confidence: 99%

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Molecular characterization of the Bidder's organ in the cane toad (Bufo marinus)

2010

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Transcriptional evolution underlying vertebrate sexual development

Valenzuela

Neuwald

Literman

2012

Developmental Dynamics

107

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Explaining the diversity of vertebrate sex-determining mechanisms ranging from genotypic (GSD) to temperature-dependent (TSD) remains a developmental and evolutionary conundrum. Using a phylogenetic framework, we explore the transcriptional evolution during gonadogenesis of several genes involved in sexual development, combining novel data from Chrysemys picta turtles (TSD) and published data from other TSD and GSD vertebrates. Our novel C. picta dataset underscores Sf1 and Wt1 as potential activators of the thermosensitive period and uncovered the first evidence of Dax1 involvement in male development in a TSD vertebrate. Contrasting transcriptional profiles revealed male-biased Wt1 expression in fish while monomorphic expression is found in tetrapods but absent in turtles. Sf1 expression appears highly labile with transitions among testicular, ovarian, and non-sex-specific gonadal formation patterns among and within lineages. Dax1's dual role in ovarian and testicular formation is found in fish and mammals but is dosage-sensitive exclusively in eutherian mammals due to its X-linkage in this group. Contrastingly, Sox9 male-biased and Aromatase female-biased expression appear ancestral and virtually conserved throughout vertebrates despite significant heterochronic changes in expression as other elements likely replaced their function in early gonadogenesis. Finally, research avenues are highlighted to further study the evolution of the regulatory network of sexual development. Developmental Dynamics 242:307-319,

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Development and steroidogenic properties of the Bidder's organ of the tadpole of Rhinella arenarum (Amphibia, Anura)

et al. 2014

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Several studies suggested that in anuran amphibians steroidogenic enzymes are critical for gonadal differentiation, proposing that the amount of sex steroids would adjust this differentiation. Among anurans, bufonids are important for the study of sex differentiation due to the presence of Bidder's organ (BO) that differentiates as a rudimentary ovary in the cephalic portion of the genital ridge. Considering that in adult males of Rhinella arenarum, the BO synthesizes estradiol, the main purpose of this work is to examine, in this species, the morphogenesis of BO and the steroidogenic capacity of this organ during larval development. BO and the proper gonads are distinguished from Gosner stage 26. During metamorphosis, BO primary oogonia develop in oogonia in nests, early previtellogenic oocytes and late previtellogenic oocytes in follicles while proper gonads remain undifferentiated. Aromatase was detected by immunohistochemistry in almost all the largest follicles of the BOs while the cytochrome P450 side-chain cleavage was observed in only few oocytes. The proper gonad was not immunoreactive in any stage. The determination of aromatase and 5α-reductase activities showed that the population of tadpoles between stages 36-41 is not homogeneous in terms of aromatase activity. In addition, from stage 26 to the end of metamorphosis, all the stages were able to produce estradiol from endogenous substrate but stages 40-41, corresponding to the end of pro-metamorphosis, produced the highest values. In conclusion, BO is able to synthesize estradiol from endogenous precursors and proper gonad remains undifferentiated at least until the end of the metamorphosis.

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Cloning and expression of candidate sexual development genes in the cane toad (Bufo marinus)

Cited by 26 publications

References 86 publications

Molecular characterization of the Bidder's organ in the cane toad (Bufo marinus)

Molecular characterization of the Bidder's organ in the cane toad (Bufo marinus)

Transcriptional evolution underlying vertebrate sexual development

Development and steroidogenic properties of the Bidder's organ of the tadpole of Rhinella arenarum (Amphibia, Anura)

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