Piprek RP et al. (2013), Sexual dimorphism of AMH, DMRT1 and RSPO1 local...

XB-ART-49232

Int J Dev Biol 2013 Jan 01;5711-12:891-5. doi: 10.1387/ijdb.130192rp.

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Sexual dimorphism of AMH, DMRT1 and RSPO1 localization in the developing gonads of six anuran species.

Piprek RP , Pecio A , Laskowska-Kaszub K , Kubiak JZ , Szymura JM .

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In vertebrates, several genes which are differentially expressed in various species, have been implicated in sex determination and gonadal differentiation. We used immunolocalization to study the expression pattern of three proteins AMH, DMRT1, RSPO1 involved in the sexual differentiation of gonads. The pattern of AMH, DMRT1 and RSPO1 expression was analyzed in X. laevis and in five other divergent anuran species: Bombina bombina, Bufo viridis, Hyla arborea, Rana arvalis and Rana temporaria during gonadal development. The pattern of expression of AMH in the developing testes of six studied anuran species was similar to that described for other vertebrates. AMH was strongly expressed in differentiating Sertoli cells. Interestingly, in B. viridis, R. arvalis and R. temporaria, AMH was also expressed in ovaries. In all studied species, DMRT1 was highly expressed in the developing testes, in both the somatic and germ cells. It was also expressed at low level in ovaries in all studied species, with the exception of H. arborea. RSPO1 was expressed in the developing ovaries, especially in the somatic cells, and was almost undetectable in developing testes in all examined anurans. These developmental expression patterns strongly suggest an involvement of AMH and DMRT1 in the development of male gonads and of RSPO1 in the female gonads. The differences in the expression patterns of these proteins in the gonads of different species might reflect the diversity of gonadal development patterns in anurans resulting from long lasting and diverged paths of their evolution.

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Species referenced: Xenopus laevis
Genes referenced: amh dmrt1 rspo1 wnt4
GO keywords: germ cell development
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	Fig. 1. Immunolocalization of AMH (A-C), DMRT1 (E-G) and RSPO1 (H-J) in the undif- ferentiated gonads, testes and ovaries in Xenopus laevis. D, control. The gonads are pointed out with arrows. o, oogonial cells; so, somatic cells; ts, testis cords. Scale bar 40 mm.
	Supplementary Fig. S1. Immunolocalization of AMH in the undifferentiated gonads, testes and ovaries in anuran amphibians. Note the overall higher level of staining in testes vs. ovaries. Rightmost column - negative control incubated with rabbit serum. The gonads are pointed with arrows, ms - cells in the gonadal mesentery, mes - mesonephros, o - oogonial cells, sg - spermatogonial cells, ts - testis cords, arrow head - gonadal medulla. Scale bar, 40 mm.
	Supplementary Fig. S2 (Left). Immunolocalization of DMRT1 in the undifferentiated gonads, testes and ovaries in anuran amphibians. Note the clearly higher level of staining in testes vs. ovaries. The gonads are pointed with arrows, mes - mesonephros, o - oogonial cells, sg - spermatogonial cells, so - somatic cells. Scale bar, 40um.
	Supplementary Fig. S3 (Right). Immunolocalization of RSPO1 in the undifferentiated gonads, testes and ovaries in anuran amphibians. Note the overall higher level of staining in ovaries vs. testes. The gonads are pointed with arrows, mes - mesonephros, o - oogonial cells, so - somatic cells, arrow head - gonadal medulla. Scale bar, 40um.