Trifold origin of the reptilian ear ossicle and its relation to the evolutionary modification of the temporal skull region.

Whereas mammals are characterized by the presence of three middle ear ossicles, reptiles have only one, the columella (stapes). Nevertheless, there is a great diversity of columellar anatomy among sauropsids, especially in the unique and cartilaginous "extracolumella"-portion. Molecular studies revealed the "columella" of chicken and quails to be formed within the second pharyngeal arch, although conflicting evidence exists for the columellar footplate and distal parts of the columella in these birds. We studied columellar development in four turtles, one lizard, and one caiman species and argue, using early blastematous stages, that, distally, the so-called "extracolumella" in turtles is mainly of quadrate, that is, first pharyngeal arch origin. Differently, the dorsal aspect of the "extracolumella" of the lizard and a part of the "dorsal columella process" of the caiman are likely quadrate-derived. This indicates only a partial homology of the distal columellar compartments among reptiles. Moreover, we observed in most species that, at early stages, the footplate differentiates from the otic capsule, which confirms widespread experimental findings of mesodermal cells contributing to the proximal part of the columella. We provide a hypothetical framework for the changes in the columella and quadrate morphology in reptilian evolution. Originally, as evidenced by the fossil record, the columella served as a stabilizing brace between the quadrate and braincase. Associated with changes in the feeding mode of late Permian taxa, the quadrate was integrated along the stress flows from biting, and in early development part of the quadrate differentiated to differently contribute to the distal part of the "columella-complex," which now contacts the tympanic membrane. In addition, part of the original otic capsule contributes to the footplate of the mobile columella, providing a connection with the inner ear.