Cell structure of developing barbs and barbules in downfeathers of the chick: Central role of barb ridge morphogenesis for the evolution of feathers.

The present ultrastructural study shows how cells organize to form the complex structure of downfeathers in chick embryos. The embryonic epidermis of the apical part of feather filaments folds inward forming barb ridges which extend toward the base of the feather. The stratification of epidermal cells in barb ridges is maintained but the basal layer loses most of the germinal activity. New cells for the growth of feather filaments are mainly produced in its basal part. In barb ridges only the original four epidermal layers of the embryonic epidermis remain to form feathers: 1) the external periderm, 2) three-five layers of the feather sheath and barb vane ridge cells, 3) subperiderm cells, and 4) basal or cylindrical cells. Periderm, sheath, barb vane ridge and cylindrical cells synthesize only alpha-keratin. Instead, cells of the subperiderm layer synthesize a small type of beta-keratin: feather beta-keratin. At hatching, the subperiderm layer is lost in most areas of the skin of the chick (apteric and scaled), and is replaced by cells containing alpha-keratin (interfollicular-apteric epidermis), scale beta-keratin (scales), beak beta-keratin (beak), and claw beta-keratin (claws). Only in feathers, cells of the original subperiderm layer remain and give origin to barb and barbule cells. The formation of separated chains of barb and barbule cells is allowed by the presence of barb vane ridge cells that function as spacers between merging cells of barb and barbule cells. Subperiderm cells elongate and merge into a syncitium to form barbules and barbs. While barbule and barb cells accumulate feather-keratin, barb vane and cylindrical cells accumulate lipids, vesicles and little alpha-keratin. These cells eventually degenerate by necrosis leaving empty spaces and lipids between barbules and barbs. No apoptosis is necessary to explain the process of carving out of barb and barbules in feathers after dissolution of the external sheath. In fact, the retraction of blood vessels nourishing the apical part of the feather filament determines anoxia and eventually necrosis of all cells of the feather. While sheath, barb vane and cylindrical cells degenerate, the keratinized syncitium forming barbs and barbules simply remain in place to form the ramifications of feathers. The formation of barb ridges is considered as the evolutionary innovation necessary for the origin of feathers. The evolution of the morphogenetic process of barb ridge formation within epidermal tubular outgrowths of the integument of ancient archosaurians was an evolutionary novelty, a true avian and theropod characteristic. Barb ridges morphogenesis determines the contemporary formation of barb and barbule cells as a unique and inseparable process so that intermediate forms of evolving feathers with only barbs but not barbules are unlikely. Barb ridges can merge with a large ridge (rachis) or into branched ridges, a process which was at the origin of the ramogenic process from which pennaceous feathers evolved.