Characterization and expression pattern of zebrafish anti-Müllerian hormone (amh) relative to sox9a, sox9b, and cyp19a1a, during gonad development

doi:10.1016/j.modgep.2005.02.008

Gene Expression Patterns

Volume 5, Issue 5, June 2005, Pages 655-667

https://doi.org/10.1016/j.modgep.2005.02.008 Get rights and content

Abstract

The role of Anti-Müllerian hormone (Amh) during gonad development has been studied extensively in mammals, but is less well understood in other vertebrates. In male mammalian embryos, Sox9 activates expression of Amh, which initiates the regression of the Müllerian ducts and inhibits the expression of aromatase (Cyp19a1), the enzyme that converts androgens to estrogens. To better understand shared features of vertebrate gonadogenesis, we cloned amh cDNA from zebrafish, characterized its genomic structure, mapped it, analyzed conserved syntenies, studied its expression pattern in embryos, larvae, juveniles, and adults, and compared it to the expression patterns of sox9a, sox9b and cyp19a1a. We found that the onset of amh expression occurred while gonads were still undifferentiated and sox9a and cyp19a1a were already expressed. In differentiated gonads of juveniles, amh showed a sexually dimorphic expression pattern. In 31 days post-fertilization juveniles, testes expressed amh and sox9a, but not cyp19a1a, while ovaries expressed cyp19a1a and sox9b, but not amh. In adult testes, amh and sox9a were expressed in presumptive Sertoli cells. In adult ovaries, amh and cyp19a1a were expressed in granulosa cells surrounding the oocytes, and sox9b was expressed in a complementary fashion in the ooplasm of oocytes. The observed expression patterns of amh, sox9a, sox9b, and cyp19a1a in zebrafish correspond to the patterns expected if their regulatory interactions have been conserved with mammals. The finding that zebrafish sox9b and sox8 were not co-expressed with amh in oocytes excludes the possibility that amh expression in zebrafish granulosa cells is directly regulated by either of these two genes.

Section snippets

Results and discussion

Mammalian embryos initially develop an indifferent gonad that has the potential of becoming an ovary or a testis (see for recent reviews (Brennan and Capel, 2004, Park and Jameson, 2005)). The Y-chromosome gene Sry stimulates early testis development (Gubbay et al., 1990, Sinclair et al., 1990) and activates the expression of the transcription factor Sox9 in Sertoli cell precursors (Sekido et al., 2004). In concert with other factors, including Sf1, Wt1, and Gata4 (Shen et al., 1994, de Santa

Cloning amh

To determine whether zebrafish possesses a gene orthologous to mammalian Amh, we searched the zebrafish genome database (Sanger Institute, Ensembl: http://www.ensembl.org/Danio_rerio) by tBLASTn comparison (Altschul et al., 1997) with the mouse AMH (NP_031471). The best zebrafish hit (ENSDART00000013803) resulting from this search, was then compared with GenBank (http://www.ncbi.nlm.nih.gov/blast/) to infer its nature, returning as best hit the Japanese eel spermatogenesis-preventing substance

Acknowledgements

We thank Joy Murphy, Amber Starks, and the University of Oregon Zebrafish Facility for providing animals, and Poh Kheng Loi and Amber Selix of the Histology Facility for sectioning. We also thank Ricard Albalat at the Department of Genetics at the University of Barcelona, for helpful comments on the manuscript. The work was supported by NIH grants R01RR10715 and HD22486 and NSF grant IBN-9728587. We thank the Spanish Ministry of Education, Culture and Sports for support for CC (EX2002–0059).

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Characterization and expression pattern of zebrafish anti-Müllerian hormone (amh) relative to sox9a, sox9b, and cyp19a1a, during gonad development

Abstract

Section snippets

Results and discussion

Cloning amh

Acknowledgements

Cell

J. Steroid. Biochem. Mol. Biol.

Cell

Dev. Biol.

Aquat. Toxicol.

Gen. Comp. Endocrinol.

Early Hum. Dev.

Mol. Cell. Endocrinol.

Trends Genet.

Biochem. Biophys. Res. Commun.

Aquat. Toxicol.

Comp. Biochem. Physiol. B Biochem. Mol. Biol.

Gene

Aquat. Toxicol.

Curr. Opin. Cell Biol.

Cell

Fed. Eur. Biochem. Soc. Lett.

J. Biol. Chem.

Dev. Biol.

Cell

J. Steroid Biochem. Mol. Biol.

Gene

Trends Genet.

J. Steroid Biochem. Mol. Biol.

J. Steroid Biochem. Mol. Biol.

J. Steroid Biochem. Mol. Biol.

Comp. Biochem. Physiol. A Mol. Integr. Physiol.

Biochem. Biophys. Res. Commun.

The transcription factor Sox9 has essential roles in successive steps of the chondrocyte differentiation pathway and is required for expression of Sox5 and Sox6

Genes Dev.

Gapped BLAST and PSI-BLAST: a new generation of protein database search programs

Nucleic Acids Res.

Banded chromosomes and the zebrafish karyotype

Zebrafish hox clusters and vertebrate genome evolution

Science

Developmental roles of pufferfish Hox clusters and genome evolution in ray-fin fish

Genome Res.

One tissue, two fates: molecular genetic events that underlie testis versus ovary development

Nat. Rev. Genet.

Functional analysis of Sox8 and Sox9 during sex determination in the mouse

Development

Two Cyp19 (P450 aromatase) genes on duplicated zebrafish chromosomes are expressed in ovary or brain

Mol. Biol. Evol.

Characterization of duplicated zebrafish cyp19 genes

J. Exp. Zool.

Genome duplication, subfunction partitioning, and lineage divergence: Sox9 in stickleback and zebrafish

Dev. Dyn.

Direct interaction of SRY-related protein SOX9 and steroidogenic factor 1 regulates transcription of the human anti-Mullerian hormone gene

Mol. Cell. Biol.

Expression and subcellular localization of SF-1, SOX9, WT1, and AMH proteins during early human testicular development

Dev. Dyn.

A quantitative and interspecific test for biological activity of anti-mullerian hormone: the fetal ovary aromatase assay

Development

Prediction of proprotein convertase cleavage sites

Protein Eng. Des. Sel.

Regulation of ovarian function: the role of anti-Mullerian hormone

Reproduction

High reproducibility of serum anti-Mullerian hormone measurements suggests a multi-staged follicular secretion and strengthens its role in the assessment of ovarian follicular status

Hum. Reprod.