Ultrastructural and immunohistochemical observations on the process of horny growth in chelonian shells

The process of growth of horny scutes of the carapace and plastron in chelonians is poorly understood. In order to address this problem, the shell of the terrestrial tortoise Testudo hermanni, the freshwater turtle Chrysemys picta, and the soft shelled turtle Trionix spiniferus were studied. The study was carried out using immunohistochemistry, electron microscopy and autoradiography following injection of tritiated histidine. The species used in the present study illustrate three different types of shell growth that occur in chelonians. In scutes of Testudo and Chrysemys, growth mainly occurs in the hinge regions by the production of cells that accumulate beta-keratin and incorporate tritiated histidine. Newly produced bundles of alpha- and beta-keratin incorporate most of the histidine. No keratohyalin is observed in the epidermis of any of the species studied here. In Testudo, newly generated corneocytes containing beta-keratin form a corneous layer to form the growing rings of scutes. In Chrysemys, newly generated corneocytes containing beta-keratin form the new, expanded corneous layer. In the latter species, at the end of the growing season (autumn/fall), thin corneocytes containing little beta-keratin are produced underneath the corneous layer, and gradually form a scission layer. In the following growing season (spring-summer) the shedding layer matures and determines the loss of the outer corneous layer. In this way, scutes expand their surface at any new molt. In Trionix, no distinct scutes and hinge regions are present and during the growing season, new corneocytes are mainly produced along the perimeter of the shell. Corneocytes of Trionix contain little beta-keratin and form a thick corneous layer in which cells resemble the alpha-layer of the softer epidermis of the limbs, tail and neck. Neither keratohyalin nor specific histidine incorporation was observed in these cells. Corneocytes are gradually lost from the epidermal surface. Dermal scutes are absent in Trionix, but the dermis is organized in 6-10 layers of plywood-patterned collagen bundles. The stratified layers gradually disappear toward the growing border of the shell. The mode of growth of horny scutes in these different species of chelonians is discussed.