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Sexually dimorphic development of mouse primordial germ cells: switching from oogenesis to spermatogenesis

Ian R. Adams and Anne McLaren*

Wellcome/CRC Institute of Cancer and Developmental Biology, Tennis Court Road, Cambridge CB2 1QR, UK



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  		Fig. 1. Development of PGCs in urogenital ridge aggregates. Donor PGCs carrying a lacZ transgene were isolated from 11.5 dpc (A,B), 12.5 dpc (C) or 13.5 dpc (D) female genital ridges, and from 11.5 dpc (E,F), 12.5 dpc (G) or 13.5 dpc (H) male genital ridges, as indicated. The donor PGCs were aggregated with male (B-E) or female (A,F,G,H) 12.5 dpc recipient urogenital ridges (UGR) and cultured for 3-4 days. Donor PGCs are identified by a cyan perinuclear dot. Donor PGCs developing as oocytes (arrowheads) have condensed chromatin staining. Donor PGCs developing as prospermatogonia (arrows) have diffuse chromatin staining and prominent nucleoli. Scale bar: 10 µm.

 


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  		Fig. 2. Representational difference analysis of 12.5 dpc PGC cDNA. RDA was used to identify cDNAs that were differentially expressed in male and female PGCs at 12.5 dpc. Five male-enriched bands were present on a gel after three rounds of RDA. The identity and size of three of these bands are shown.

 


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  		Fig. 3. Expression of Ptgds in embryonic gonads. Whole-mount in situ hybridisation for Ptgds was performed on isolated 11.5 dpc (A,B), 12.5 dpc (C) or 13.5 dpc (D) male and female urogenital ridges, as indicated. We/We testes (14.5 dpc) were used in (E). At 11.5 dpc, paired urogenital ridges with intervening mesenchyme are shown with the genital ridges lying on top of the mesonephroi along the left and right edges of the tissue. At 12.5-14.5 dpc, single urogenital ridges are shown, orientated with the mesonephros on the left and the genital ridge on the right. Sections of 13.5 dpc male urogenital ridges after whole-mount in situ hybridisation for Ptgds are shown in (F). Sertoli cells are marked with arrowheads, prospermatogonia with arrows. Sections are stained with neutral red to show the nuclei. Transcript expression is visualised by a dark blue/purple precipitate. Scale bars: in E, 200 mm in A-E; in F, 10 µm.

 


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  		Fig. 4. Exogenous prostaglandin D2 masculinises female urogenital ridges in culture. Female urogenital ridges were cultured with or without exogenous prostaglandin D2 in the culture medium, fixed in Bouin’s solution and sections stained with Haematoxylin and Eosin. (A-D) Sections are oriented with the mesonephros (m) on the left and the genital ridge (g) on the right. (A) Control female urogenital ridge cultured without prostaglandin D2. (B-D) Female urogenital ridges cultured with prostaglandin D2. (B) Disorganised cord-like structures in the masculinised region of the genital ridge (), and the ovarian region () of the genital ridge. (C) Eosinophilic cells bulging from the genital ridge into the mesonephros (arrow). (D) The mass of gonadal eosinophilic cells is located along the boundary with the mesonephros (arrow). (E-G) Higher magnification of female urogenital ridges cultured with prostaglandin D2. Thick arrows labelled PSG indicate prospermatogonia, peritubular myoid-like cells are indicated with small arrows and Sertoli-like cells are indicated with arrowheads. (E) A disorganised cord in the masculinised region of the urogenital ridge shown in B. (F,G) Sections located at the edge of the bulge of eosinophilic cells spilling into the mesonephros, from the urogenital ridge shown in C. Scale bars: 50 µm in A-D; 10 µm in E-G.

 


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  		Fig. 5. Exogenous prostaglandin D2 induces expression of AMH in cultured female urogenital ridges. Contralateral urogenital ridges from a female 11.5 dpc embryo were cultured without prostaglandin D2 (A), or with prostaglandin D2 (B). AMH expression was then examined by immunohistochemistry. Positive signal is shown by the brown precipitate, nuclei were counterstained blue with Haematoxylin. Images are from the genital ridge regions. Scale bar: 50 µm.

 


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  		Fig. 6. The role of prostaglandin D2 signalling in testis development. (A) In a XX {leftrightarrow} XY chimaeric testis, a mixture of XX and XY supporting cells (ovoid cells) can be in contact with a bipotential primordial germ cell (outlined in red). XY-supporting cells express Sry (blue), and differentiate into pre-Sertoli cells. The pre-Sertoli cell produces the uncharacterised masculinising signal (blue arrow, ?) that acts on PGCs directing them to develop as prospermatogonia. The pre-Sertoli cell also expresses prostaglandin D synthase (purple). Prostaglandin D2 acts as a local paracrine factor to induce nearby XX supporting cells to differentiate into pre-Sertoli cells (purple arrow, PD2). Commitment of PGCs to prospermatogonial development results in the prospermatogonia initiating expression of prostaglandin D synthase (purple). Prostaglandin D2 from the prospermatogonia induces nearby XX supporting cells to differentiate into pre-Sertoli cells (purple arrow). (B) In XY mismatch testis with ‘weak’ Sry expression and an early acting programme for ovarian differentiation, some XY supporting cells express Sry effectively (blue), while others do not (white). Testis development proceeds similarly to the chimaeric testis, with prostaglandin D2 inducing undifferentiated supporting cells to differentiate into pre-Sertoli cells. (C) In XY testis with ‘strong’ Sry expression, Sry is expressed effectively in all the supporting cells (blue), and the paracrine prostaglandin D2 signal for inducing pre-Sertoli cell differentiation is somewhat redundant.

 

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© The Company of Biologists Ltd 2002