Computational Approaches and Observer Variation in the 3D Musculoskeletal Modeling of the Heads of Anolis.

High-resolution imaging, 3D modeling, and quantitative analyses are equipping evolutionary biologists with new approaches to understanding the variation and evolution of the musculoskeletal system. However, challenges with interpreting DiceCT data and higher order use of modeled muscles have not yet been fully explored, and the error in and accuracy of some digital methods remain unclear. West Indian Anolis lizards are a model clade for exploring patterns in functional adaptation, ecomorphology, and sexual size dimorphism in vertebrates. These lizards possess numerous jaw muscles with potentially different anatomies that sculpt the adductor chamber of the skull. Here we test approaches to quantifying the musculoskeletal shape of the heads of two species of Anolis: A. pulchellus and A. sagrei. We employ comparative approaches such as DiceCT segmentation of jaw muscles, 3D surface attachment mapping, and 3D landmarking with the aim of exploring muscle volumes, 3D muscle fiber architecture, and sexual dimorphism of the skull. We then compare sources of measurement error in these 3D analyses while also presenting new 3D musculoskeletal data from the Anolis feeding apparatus. These findings demonstrate the accessibility and repeatability of these emerging techniques as well as provide details regarding the musculoskeletal anatomy of the heads of A. pulchellus and A. sagrei which show potential for further research of comparative biomechanics and evolution in the clade.

3D Muscle Architecture of the Pectoral Muscles of European Starling (Sturnus vulgaris).

Avian flight is achieved through a number of modifications to the body, including the pectoral girdle, yet little is known about the architecture of the pectoral musculature. Muscle architecture is a critical variable in determining the biomechanical function of the vertebrate musculoskeletal system; however, accurate three-dimensional (3D) understanding of muscle architecture has been historically difficult to acquire. Here, we present a musculoskeletal model of a European starling (Sturnus vulgaris) pectoral girdle generated from iodine contrast-enhanced micro-computed-tomography (CT) data and 3D fiber tracking analysis. We used a template-based fiber-tracking algorithm to reconstruct muscle fibers in 3D based on grayscale differences in CT images, which allowed us to estimate fascicle lengths, pennation angles, muscle volumes, and physiological cross-sectional area. Our modeled muscles were qualitatively accurate; however, quantitative muscle architecture data differed between digital and traditional gross-dissection methods reflecting the complex organization of the tissue and differing natures of data collection. We found that model quality is affected by the resolution of CT image data and the fiber-tracking program's input parameters. Nonetheless, digital fiber tracking offers numerous advantages over gross-dissection methods, most importantly, the ability to visualize and quantify entire muscles in three-dimensions, yielding a much more accurate estimation of whole muscle architecture.