Expression and Functional Analysis of Dkk1 during Early Gonadal Development

Combes, Alexander N.; Bowles, Josephine; Feng, Chunliang; Chiu, H.S.; Khoo, Poh-Lynn; Jackson, Andrew; Little, Melissa H.; Tam, Patrick; Koopman, Peter

doi:10.1159/000327709

Cited by 16 publications

(14 citation statements)

References 81 publications

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“…As such, both DKK1 and DACT1 enable the axin/ Gsk3/APC complex to degrade β-catenin and minimize the expression of β-catenin/LEF/TCF-responsive genes, resulting in a different cell type than those lacking either DKK1 or DACT1. DKK1 has been found to play a role in normal development of testis in mice Combes et al, 2011]. But unlike mice, we found that dkk1 was upregulated in ovaries rather than testes.…”

Section: Discussioncontrasting

confidence: 63%

Antagonists to the Wnt Cascade Exhibit Sex-Specific Expression in Gonads of Sexually Mature Shovelnose Sturgeon

Amberg

Goforth

Sepúlveda³

2013

Sex Dev

343

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Little is known regarding molecular mechanisms involved in sex determination and differentiation in sturgeon species. We addressed this knowledge gap by using next generation pyrosequencing technology to provide transcript libraries and species-specific sequences for mature gonads of shovelnose sturgeon, Scaphirhynchus platorynchus. We then mined these libraries to identify gender-specific transcripts and quantified relative transcript abundance using quantitative real-time polymerase chain reaction (qPCR). We detected a limited number of genes known to play a role in sex differentiation in other species. The sequence for dmrt1 was found only in the testes library. The abundance of dmrt1 differed slightly between the sexes, but the melt curve suggests that there may be different isoforms of dmrt1 in ovaries and testes of shovelnose sturgeon. The transcription factor foxl2 was 5.3-fold greater in ovaries than in testes. Two antagonists to the Wnt cascade, dickkopf-1 (dkk1) and dapper-1 (dact1), were found only in the ovary library. Results from qPCR indicated that dkk1 and dact1 were upregulated 1,819.1- and 207.5-fold, respectively, in ovaries compared with testes. These results suggest that antagonists to the Wnt cascade may play significant roles in sex differentiation and gonadal development in sturgeon and could serve as sex markers in this group of ancient fish.

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Section: Discussioncontrasting

confidence: 63%

Antagonists to the Wnt Cascade Exhibit Sex-Specific Expression in Gonads of Sexually Mature Shovelnose Sturgeon

Amberg

Goforth

Sepúlveda³

2013

Sex Dev

343

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“…qRT-PCR is commonly used to study gene expression during gonadogenesis (Abramyan et al 2009;Barrionuevo et al 2006;Barske and Capel 2010;Beverdam and Koopman 2006;Bogani et al 2009;Bouma et al 2005Bouma et al , 2007Bradford et al 2009;Britt et al 2002;Chang et al 2008;Chassot et al 2008;Chen et al 2012;Childs et al 2011;Combes et al 2011;Cupp et al 2003;DeFalco et al 2014;Hiramatsu et al 2009;Houmard et al 2009;Koubova et al 2006;Li et al 2012;Liu et al 2015;Maatouk et al 2008;Manuylov et al 2007;Molyneaux et al 2003;Moniot et al 2009;Munger et al 2013;Nef et al 2003Nef et al , 2005Nicol and Yao 2015;Parma et al 2006;Rolland et al 2011;Ross et al 2009;Smith et al 2008aSmith et al , b, 2009Tang et al 2008;Tevosian et al 2002;Wilhelm et al 2005). In the majority of studies of gonad development, primers labeled with SYBR Green fluorochrome are used although TaqMan probes have also been applied Liu et al 2015;Smith et al 2008a, b).…”

Section: Pcr-based Methods Of Gene Expression Analysismentioning

confidence: 99%

“…The first report of detection of DNA:RNA hybridization was given by Gall and Pardue (1969). This method is frequently used in gonadal development studies in both the post-embedding version (ISH) (Abramyan et al 2009;Best et al 2008;Bradford et al 2009;Chassot et al 2008;El Jamil et al 2008;Hanley et al 2000;Kim et al 2006;Maatouk et al 2008;Martineau et al 1997;Matzouk et al 1995;Moniot et al 2009;Morais da Silva et al 1996;Nef et al 2003;Oulad-Abdelghani et al 1996;Parma et al 2006;Schmahl et al 2004;Sekido et al 2004;Wilhelm et al 2005) and pre-embedding or whole mount-ISH (WM-ISH or WISH) (Barrionuevo et al 2006;Bishop et al 2000;Bogani et al 2009;Combes et al 2009aCombes et al ,2011Coveney et al 2008;Jameson et al 2012;Koubova et al 2006;Manuylov et al 2007;Matsuyama et al 2005;Nef et al 2005;Rolland et al 2011;Smith et al 2008a, b;Stebler et al 2004). The core idea in the ISH technique is the hybridization of a labeled complementary probe to mRNA allowing the identification of a specific gene transcript under a microscope.…”

Section: Identification Of Mrna: In Situ Hybridizationmentioning

confidence: 99%

“…DNA microarray techniques require RNA isolation and its conversion into cDNA through reverse transcription; cDNA is labeled with fluorochrome, then applied onto chips, and hybridized to probes (cDNA or oligonucleotides) previously attached to the chip; after rinsing, only labeled sequences that bound complementary probes remain on the chip and signals emitted by them are detected. In gonadogenesis studies, microarrays were used for gene expression analysis of RNA isolated from whole developing gonads and thus from many different cell lines (Combes et al 2011;Coveney et al 2008;Gallardo et al 2007;Garcia-Ortiz et al 2009;Grimmond et al 2000;Houmard et al 2009;Liu et al 2015;Munger et al 2013;Nicol and Yao 2015;Ottolenghi et al 2007;Rolland et al 2011;Small et al 2005), whereas other microarray studies were conducted on particular gonadal cell lines, e.g., on germ cells (Jameson et al 2012;Molyneaux et al 2003;Pan et al 2005), pre-Sertoli and pre-granulosa cells (Beverdam and Koopman 2006;Bouma et al 2007;Jameson et al 2012;Nef et al 2005), epithelial cells of gonadal primordium (Hummitzsch et al 2013;Kusaka et al 2010), and interstitial and endothelial cells (Jameson et al 2012). The microarray technique was also used to identify noncoding RNAs in developing gonads (Chen et al 2012). 14.14.6 RNA Sequencing RNA sequencing (RNA-Seq) is one of the latest high-throughput methods used for gene expression analysis (from 2008).…”

Section: Microarraymentioning

confidence: 99%

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Methods for the Study of Gonadal Development

Piprek

2016

Results and Problems in Cell Differentiation

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Current knowledge on gonadal development and sex determination is the product of many decades of research involving a variety of scientific methods from different biological disciplines such as histology, genetics, biochemistry, and molecular biology. The earliest embryological investigations, followed by the invention of microscopy and staining methods, were based on histological examinations. The most robust development of histological staining techniques occurred in the second half of the nineteenth century and resulted in structural descriptions of gonadogenesis. These first studies on gonadal development were conducted on domesticated animals; however, currently the mouse is the most extensively studied species. The next key point in the study of gonadogenesis was the advancement of methods allowing for the in vitro culture of fetal gonads. For instance, this led to the description of the origin of cell lines forming the gonads. Protein detection using antibodies and immunolabeling methods and the use of reporter genes were also invaluable for developmental studies, enabling the visualization of the formation of gonadal structure. Recently, genetic and molecular biology techniques, especially gene expression analysis, have revolutionized studies on gonadogenesis and have provided insight into the molecular mechanisms that govern this process. The successive invention of new methods is reflected in the progress of research on gonadal development.

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“…Dkk1 (Dickkopf homologue 1) served as a control as it is enriched in the embryonic testis [37,38] . It was in fact robustly expressed in the WT embryonic testes, not detected in the WT ovaries, and reduced in the Wnt-4-deficient testes ( Fig.…”

Section: Wnt-4 Signalling Also Controls the Genes Associated With Thementioning

confidence: 99%

Identification of the genes regulated by Wnt-4, a critical signal for commitment of the ovary

Naillat

Yan

Karjalainen

et al. 2015

Experimental Cell Research

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The indifferent mammalian embryonic gonad generates an ovary or testis, but the factors involved are still poorly known. The Wnt-4 signal represents one critical female determinant, since its absence leads to partial female-to-male sex reversal in mouse, but its signalling is as well implicated in the testis development. We used the Wnt-4 deficient mouse as a model to identify candidate gonadogenesis genes, and found that the Notum, Phlda2, Runx-1 and Msx1 genes are typical of the wild-type ovary and the Osr2, Dach2, Pitx2 and Tacr3 genes of the testis. Strikingly, the expression of these latter genes becomes reversed in the Wnt-4 knock-out ovary, suggesting a role in ovarian development. We identified the transcription factor Runx-1 as a Wnt-4 signalling target gene, since it is expressed in the ovary and is reduced upon Wnt-4 knock-out. Consistent with this, introduction of the Wnt-4 signal into early ovary cells ex vivo induces Runx-1 expression, while conversely Wnt-4 expression is down-regulated in the absence of Runx-1. We conclude that the Runx-1 gene can be a Wnt-4 signalling target, and that Runx-1 and Wnt-4 are mutually interdependent in their expression. The changes in gene expression due to the absence of Wnt-4 in gonads reflect the sexually dimorphic role of this signal and its complex gene network in mammalian gonad development.

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Expression and Functional Analysis of Dkk1 during Early Gonadal Development

Cited by 16 publications

References 81 publications

Antagonists to the Wnt Cascade Exhibit Sex-Specific Expression in Gonads of Sexually Mature Shovelnose Sturgeon

Antagonists to the Wnt Cascade Exhibit Sex-Specific Expression in Gonads of Sexually Mature Shovelnose Sturgeon

Methods for the Study of Gonadal Development

Identification of the genes regulated by Wnt-4, a critical signal for commitment of the ovary

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