Describing the musculature of mystacial pads in harbour seals (Phoca vitulina) using diceCT.

Pinnipeds have long, sensitive, moveable mystacial vibrissae. In other mammals, intrinsic muscles contribute to protracting the vibrissae. However, the mystacial muscles of pinnipeds have not yet been systematically described. Using traditional histological methods provides us with two-dimensional muscle images, but having the ability to visualise these structures in three dimensions would allow for a more comprehensive understanding of pinniped vibrissal anatomy, especially given the challenges posed by their large and extremely curved mystacial pad. We predicted that harbour seals would have large, regular intrinsic muscles due to their well-organised, moveable vibrissae. We adopted diffusible iodine contrast-enhanced computer tomography (diceCT) to describe, for the first time, the three-dimensional architecture of the mystacial vibrissal muscles found in harbour seals. Our observations show that their vibrissae are organised into grids within the mystacial pad. We identified both sling-shaped and oblique intrinsic muscles that connect one vibrissae to the next in the same row. We also identified extrinsic muscles, including the m. nasolabialis, m. maxillolabialis, m. levator nasolabialis and m. orbicularis oris. Contrary to our prediction, the intrinsic muscles were not very large, although they were regularly distributed throughout the pad. Rather, the extrinsic muscles, particularly the m. nasolabialis and m. maxillolabialis were large, deep and well-defined, running throughout the length of the mystacial pad. Therefore, we suggest that these extrinsic muscles, the m. nasolabialis and m. maxillolabialis, are responsible for driving vibrissal protraction underwater. These findings demonstrate the importance of three-dimensional visualisation techniques in advancing our understanding of mystacial anatomy and function in pinnipeds.