Development of the malleus and incus from Meckel's cartilage in the mouse. (A, B) Alcian blue‐stained section of Meckel's cartilage at E13.5 (A) and E14.5 (B). (C) Expression of the joint marker, Gdf5 on a sagittal section at E15.5, showing expression in the joint region between the malleus and incus. (D, E) Alcian blue‐ and Alizarin red‐stained skeletal preparations of the caudal end of Meckel's cartilage, showing the connection to the malleus at P0 (D) and its separation at P3 (E). (F) Schematic representing the separation of the ossicles, between E13.5 and P3. At E13.5 (A, F), the malleus and incus are still fused to the caudal end of Meckel's cartilage. At E14.5 (B, C, F), the malleus and incus have separated (arrow), and the joint marker Gdf5 is expressed in the joint region between the malleus and incus at E15.5 (arrow). At P0 (D, F), Meckel's cartilage is still fused to the malleus (arrow). At P3 (E, F), the malleus has separated from Meckel's cartilage (arrow). I, incus; M, malleus; mk, Meckel's cartilage; S, stapes. (A–C taken from Amin & Tucker, , © 2006 Wiley‐Liss, Inc; D–E taken from Anthwal et al. , © 2012 The Authors. Journal of Anatomy © 2012 Anatomical Society).