PMC:523229 / 17007-20551 JSONTXT

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The Gdf5-Cre/Bmpr1afloxP conditional knockout mice were viable and survived to adulthood, showing that the Gdf5-Cre driver can bypass the early embryonic lethality previously reported in animals with a null mutation in the Bmpr1a locus (Mishina et al. 1995).\nFigure 3 Gdf5-Cre-Mediated Deletion of Bmpr1a\n(A) Breeding strategy simultaneously deletes Bmpr1afloxP and allows visualization of Gdf5-Cre-mediated recombination by lacZ expression from R26R.\n(B–E) 5-week-old mutant and control mice stained with Alcian blue to mark cartilage and alizarin red to mark bone. (B) Ankle of control with strong blue staining lining each joint (arrowheads). (C) Ankle of mutant showing an absence of blue staining in most regions (arrowheads) and a joint fusion between the central (c) and second (2) tarsals (arrow). (D) Control and (E) mutant metatarsal/phalangeal joint which lacks blue staining in articular regions (arrowheads) but retains staining in the growth plate (asterisks).\n(F) Control forelimb.\n(G) Mutant forelimb with webbing between the first and second digit (black arrowhead). The viable Gdf5-Cre/Bmpr1afloxP mice showed several phenotypes. First, the conditional knockout mice had shorter ears that often lay flatter against their heads than controls (controls 13.1 ± 0.1 mm long, n = 38; mutants 11.8 ± 0.2 mm, n = 11; p \u003c 0.0001). BMP signaling is known to be required for growth of the external ear of mice (Kingsley et al. 1992), and this phenotype likely reflects loss of Bmpr1a function in the fraction of ear cells that express the Gdf5-Cre transgene. Most mutant mice also showed soft tissue syndactyly or retention of webbing between the first and second digits of their feet, a phenotype that was more frequent and more severe in the forelimbs (201 of 220, or 91%, of forefeet and 109 of 220, or 50%, of hindfeet). Finally, mutant animals showed obvious skeletal changes in whole-mount skeletal preparations. At some sites in the ankles, joints seemed to be missing entirely, with fusion of bones that would normally be separate. For example, the second distal tarsal was fused to the central tarsal bone in every conditional knockout animal examined (18 of 18), a phenotype not observed in controls (zero of 18) (Figure 3B and 3C). At other locations, joints had clearly formed but showed dramatic loss of staining with the cartilage matrix marker Alcian blue (Figure 3B–3E) (unpublished data). Normal Alcian blue staining was seen in non-articular regions, such as the cartilaginous growth plate (Figure 3D and 3E, asterisk). These data suggest that Bmpr1a function is required for the formation of specific joints in the ankle region and for either generation or maintenance of articular cartilage in most other joints of the 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