Fig. 3. PGC number is progressively impaired in Pin1^-/- embryos. (A,C,E,G) Wild-type. (B,D,F,H) Pin1^-/-. (A,B) Light micrograph images of 8.5 dpc wild-type (A) and Pin1^-/- (B) wholemount embryos after alkaline phosphatase assays show PGCs (arrow) are properly allocated at the base of the allantois. (C,D) Light micrograph images of 9.5 dpc wild-type (C) and Pin1^-/- (D) wholemount embryos after alkaline phosphatase assays show PGCs (arrows) migrating through the hindgut. The Pin1^-/- embryo has fewer PGCs. (E-H) Confocal images of 13.5 dpc wild-type (E,G) and Pin1^-/- (F,H) male XY (E,F) and female XX (G,H) gonads after whole-mount immunohistochemistry. PGCs (red, round cells, white arrows) are identified as large round cells by their surface antigen PECAM. Testis cords are identified by laminin at the basement membrane as green crescents (white arrowheads). The Pin1-deficient gonads have few PGCs, but normal cords are formed in the male gonad. (I) PGC number in male XY (left, blue) and female XX (right, pink) whole embryos (8.5 and 9.5 dpc) and comparable gonadal sections (11.5-13.5 dpc) at different stages: wild-type, solid; Pin1^-/-, hatched. Values represent mean±s.e.m. *Statistically significant (ANOVA): not significant at 8.5 dpc; P<0.0005 at 9.5 dpc; P<0.0001 at 11.5-13.5 dpc. Subtests at each age were legitimatised by the genotype x age interaction, P<0.0001.

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