Protein synthesis in tissues cultured from the bovine hoof

The mechanical strength of the bovine hoof depends on keratinization of cells in the germinal layers of the epidermis. Histological examination of hoof tissues in calves and young heifers has identified disturbances in this keratinization process which would result in ineffective hoof development and could precipitate lameness. Short-term culture of bovine hoof tissue was used to investigate epidermal keratinization. Cell function remains viable in these cultures. The rate of protein synthesis, measured by [35S]-methionine incorporation, continued for at least 3 h in culture. Radiolabelled proteins in tissue homogenates were separated by SDS-polyacrylamide gel electrophoresis and characterised by fluorography and were representative of the proteins found in hoof tissue. Three prominent radiolabelled bands were identified as keratins and actin by Western blotting. Immunohistochemistry showed that keratin was localised principally in the epidermal layers, and microautoradiography indicated that this was the major site of protein synthesis. Hoof tissues cultured under these conditions provide a useful system for studying the acute regulation of epidermal keratinization.