Gonadal differentiation, sex determination and normal Sry expression in mice require direct interaction between transcription partners GATA4 and FOG2
Sergei G. Tevosian1,*,
Kenneth H. Albrecht2,
John D. Crispino3,
Yuko Fujiwara1,
Eva M. Eicher2 and
Stuart H. Orkin1,
1 Division of Hematology and Oncology, Children’s Hospital and Dana Farber Cancer Institute, Department of Pediatric Oncology, and Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, USA
2 The Jackson Laboratory, Bar Harbor, ME, USA
3 The Ben May Institute for Cancer Research, University of Chicago, Chicago, IL, USA
* Present address: Department of Genetics, Dartmouth Medical School, Hanover, NH, USA
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Fig. 1. E17.5 Fog2–/–  MHC-Fog2 transgenic and normal mouse gonads. (A-C) Macroscopic view of the urogenital systems in male (A), female (B) and XY mutant (C) E17.5 fetuses. Mutant XY gonads are severely reduced in size and have not descended. a, adrenal, b, bladder, g, gonad, k, kidney. Only one paired organ is labeled. (D,E) Representative transverse sections through gonads at the same magnification of (D) normal testis and (E) Fog2–/– undifferentiated XY gonads (arrowhead). Notice the lack of testis cord development and greatly reduced size of mutant gonad.
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Fig. 2. Expression of molecular markers of early gonadal development. (A-D) In situ hybridization of E12.5 XY gonads with Sf1 RNA probe. (A,C) Normal controls. (B) Fog2–/–. (D) Gata4ki/ki. Gonad (red arrow) and mesonephros (arrowhead) are indicated. (E-J) Analysis of SF1 (red) and WT1 (green) protein expression in E13.25 control XY gonads (E,H), control XX ovary (F,I) and XY Fog2–/– gonad (G,J) using immunofluorescent histochemistry and confocal microscopy. Notice that the XY Fog2–/– gonad lacks testis cords and resembles the control XX ovary at this stage of development.
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Fig. 3. Analysis of gonad differentiation using immunofluorescent histochemistry and confocal microscopy. (A-C) E12.5 Gata4ki/ki gonads at the time when testis cords are beginning to develop in normal XY gonads. (D-F) E13.5 XY Fog2–/– gonads at the time when testis cord are normally well developed and surrounded by a basal lamina in control XY gonads. (A,D) XX control gonads, (B,E) XY control gonads and (C,F) mutant XY gonads. Laminin, green; GATA4, red; PECAM, blue. In each image the gonad is above the mesonephros.
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Fig. 4. Sry expression. Semi-quantitative RT-PCR analysis of Sry expression in E11.5 (15-20 tail somite stage) XY gonads. A representative analysis of Sry expression relative to Lhx1 expression in the gonads of Fog2–/–, Fog2+/– and Fog2+/+ fetuses at the 18 tail somite stage is shown.
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Fig. 5. Whole-mount in situ hybridization analysis of Sertoli cell marker gene expression in control and mutant E13.5 XY gonads. Left panels: Dhh (top), Sox9 (middle) and Mis (bottom). Right panels: Sox9 (top) and Mis (bottom). g, gonad; m, mesonephros. None of the Sertoli cell markers examined is expressed in mutant gonads.
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Fig. 6. Whole-mount in situ hybridization analysis of Leydig cell markers (encoding steroidogenic enzymes) and the Wnt4 gene in E13.5-14.5 gonads. (A,C,E) Control gonads/mesonephroi; (B,D,F), Fog2–/– gonads/mesonephroi. Steroidogenic genes are not expressed in the absence of Fog2 (B,D,F), indicating that Leydig cell differentiation is blocked. Wnt4 expression is downregulated in XY normal gonads (G), but persists in XX control (H) and XY Fog2–/– gonads (I).
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© The Company of Biologists Ltd 2002
