Reevaluating Musculoskeletal Linkages in Suction-Feeding Fishes with X-Ray Reconstruction of Moving Morphology (XROMM).

Suction-feeding fishes encompass a vast diversity of morphologies and ecologies, but during feeding they all rely on musculoskeletal linkages and levers to transform the shortening of muscle into 3D expansion of the mouth cavity. To relate the shape of these skeletal elements to their function in expansion of the mouth, four-bar linkage models have been developed and widely used in studies of ecology, evolution, and development. However, we have lacked the ability to test the predictions of these 2D linkage models against the actual 3D motions of fishes' skulls. A new imaging method, X-ray Reconstruction of Moving Morphology (XROMM), now makes it possible to measure 3D skeletal motions relative to other bones within the head and relative to the fish's body, and thereby to examine directly the proposed linkages. We used XROMM to examine the opercular linkage, in which shortening of the levator operculi muscle is hypothesized to retract the operculum, and thereby the interoperculum and interoperculomandibular ligament to generate depression of the lower jaw about the quadratomandibular joint. XROMM animations of suction strikes in largemouth bass revealed that the operculum is indeed retracted relative to the suspensorium as the levator operculi muscle shortens and the jaw depresses. However, the four-bar model of this linkage overestimates the depression of the jaw by nearly a factor of two. Therefore, caution should be used in interpreting and applying the predictions of this linkage model. When we measured kinematics relative to the fish's body, we found that the operculum was relatively stable, whereas the suspensorium was elevated along with the neurocranium, pushing the quadratomandibular joint forward to produce depression of the jaw. Thus, it is the epaxial muscles elevating the neurocranium that powers depression of the jaw through the opercular linkage. However, the levator operculi muscle plays a crucial role in stabilizing the operculum to allow elevation of the head to produce depression of the lower jaw. These results support the role of cranial muscles in controlling and transmitting power from the axial muscles, rather than generating substantial power themselves. We also demonstrate the utility of XROMM for assessing the function of this, and other, cranial linkages in suction-feeding fishes.