Mammalian development does not recapitulate suspected key transformations in the evolution of the mammalian middle ear

The tympanic ring, malleus and incus of the mammalian middle ear (MME) derive from the ancestral primary jaw joint of land vertebrates. In Mesozoic mammals, evolutionary detachment of the MME from the lower jaw occurred when Meckel’s cartilage - the last connection between MME and dentary – disappeared. This disappearance is famously recapitulated in early mammalian development. Further developmental recapitulation of Mesozoic MME detachment is thought to occur in the form of negative allometry and posterior/medial replacement of MME bones relative to the jaw joint. However, despite being widely accepted, such detailed recapitulation scenarios have never been quantified. Here we show, based on µCT scans of developmental series of several marsupials and monotremes, that negative allometry of MME bones relative to the skull occurs only after MME detachment, ruling it out as a developmental detachment trigger; additionally, there is no positional change of ectotympanic or malleus relative to the dentary. Differential positioning of MME bones in the two monotreme species is also not developmentally recapitulated. Our results challenge the developmental prerequisites of widely accepted evolutionary scenarios regarding MME detachment. Rather, we observe an association of MME detachment and dental eruption, suggesting a detachment trigger relating to the onset of dentary function.