PMC:523229 / 22493-25408 JSONTXT

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At this stage, the Gdf5 gene is normally expressed in stripes that mark the sites of joint formation (Figure 4F), and the gene for the major collagen protein of cartilage matrix (Col2a1) is down-regulated in the interzone region (Figure 4E). In contrast, Col2a1 staining extended completely through the joint region between the second and central tarsal of Gdf5-Cre/Bmpr1afloxP mutants (Figure 4H, black arrow), and Gdf5 expression was seen only as a small notch extending into where the joint should be forming (Figure 4I, bracket). These data suggest that the fusions seen between ankle bones in postnatal mutant skeletons are the result of incomplete segmentation of skeletal precursors during embryonic development, a defect confined to some locations in the ankle.\nFigure 4 Bmpr1a Is Expressed in Joints and Is Required for Continued Joint Formation in the Ankle Region\n(A) Diagram of ankle bones from a wild-type mouse; bones fusing in mutant are colored red. Roman numerals II–IV, metatarsals; 2, 3, and 4/5, distal row of tarsal bones; c, central tarsal bone; ta, talus; ca, calcaneus.\n(B and C) In situ hybridization at E15.5 showing that Bmpr1a is expressed in ankle joint interzones (B, arrowheads) and in the forming articular regions of the phalangeal joints (C, arrowheads).\n(D) Near adjacent section to (C) showing Gdf5-Cre induced LACZ expression from R26R in the forming joints of the digits (arrowheads).\n(E–J) Marker gene expression and R26R LACZ staining patterns on near adjacent sections of control and mutant embryos. In control mice at E15.5 ankle joints are clearly delineated as regions that have down-regulated Col2 (E), express Gdf5 throughout (F), and express LACZ in most cells (G; white arrowheads and black arrows). In mutant embryos at the same stage, joint formation is incomplete. Faint Col2 expression can be seen connecting a medial region of tarsal 2 with metatarsal II (H, white arrowhead), and Gdf5 expression does not extend all the way across the joint at this location (I, white arrowhead). Between tarsals c and 2, mutants express Col2 across the normal joint-forming region (H, black arrow) and lack expression of Gdf5 at sites where skeletal fusions are observed (I, black arrow and bracket). 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