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. 2016 Jun 22;6:28405. doi: 10.1038/srep28405

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Copyright © 2016, Macmillan Publishers Limited

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(A) Phenotypic analysis of Hand2^CAT/+; Wnt1-Cre (Hand2^NC) mice. (a,b) Facial appearance of postnatal day (P) 1 wild-type (a) and Hand2^NC (b) mice. Hand2^NC mutants (b) exhibit brachycephaly, eyelid colobomas (arrowhead in b), and small pinna (arrow in b). (c–l) Alizarin red (mineralised bone) and alcian blue (cartilage) staining of P1 wild-type (c,e,g,i,k) and Hand2^NC (d,f,h,j,l) mice. Hand2^NC mutants demonstrate the transformation of the maxilla (mx) into a duplicated mandibular bone (mb*), which appears as a mirror image of the mandibular bone (mb). Hypoplasia of the frontal bones (fb) and frontal foramina (asterisk) are observed in the skull vault of Hand2^NC mutants (l). (B) Phenotypic analysis of Hand1^CAT/+; Wnt1-Cre (Hand1^NC) mice. (a,b) Facial appearance of P1 wild-type (a) and Hand1^NC (b) mice. Hand1^NC mutants (b) exhibit brachycephaly, eyelid colobomas (arrowhead in b), and small pinna (arrow in b). (c–l) Bone staining of P1 wild-type (c,e,g,i,k) and Hand1^NC (d,f,h,j,l) mice. Hand1^NC mutants have a partially duplicated mandible (mb*). Hypoplasia of the frontal bones (fb) and frontal foramina (asterisk) are observed in the skull vault of Hand1^NC mutants (l). (C) The middle ear phenotype of P1 wild-type (a) and Hand2^NC (b) mice. In wild-type mice (a), the malleus (ma), incus (in), stapes (st), gonial bone (gn), tympanic ring (tr), and Meckel’s cartilage (mc) are observed. In Hand2^NC mutants, the malleus, gonial bone, and tympanic ring are absent or hypomorphic. (D) The hyoid bone phenotype of P1 wild-type (a) and Hand2^NC (b) mice. The ossification of the hyoid bones is hypoplastic in Hand2^NC mutants. mx, maxilla; mb*, duplicated mandibular bone; mb, mandibular bone; fb, frontal bone; tr, tympanic ring; lo, lamina obturans; jg, jugal bones; tb, temporal bone; bs, basisphenoid bone; ui, upper incisors; li, lower incisors; li*, duplicated lower incisors; bo, basioccipital bone; pmx, premaxilla; hb, hyoid bone; pt, pterygoid; ppmx, palatal process of maxilla; pppl, palatal process of palatine; agp, angular process; crp, coronoid process; cdp, condylar process; pb, parietal bone; vo, vomer; hy, hyoid bone, with lesser (lh) and greater (gh) horns; tc, thyroid cartilage.