A simple and non-invasive method for analyzing local immune responses in vivo using fish fin

Immunocompetence is an important parameter that reflects disease resistance in fish. Very few methods to examine immunocompetence in vivo have been developed, even for mammals. In the present study, we present a unique method for analyzing local immune responses using fish fin. We first demonstrated the migration of granulocytes to the site of zymosan injection in fin in a dose-dependent manner and that this could be easily observed macroscopically due to the fin membrane transparency. We also demonstrated phagocytic activity of accumulated leukocytes after zymosan administration and that almost all phagocytes were granulocytes. In addition, we succeeded to detect respiratory burst activity of granulocytes in vivo without any in vitro treatment of cells, indicating that our present method is suitable for the analysis of granulocyte phagocytic function in vivo. The method provides a unique tool applicable for fishes that possess transparent fins and may lead to better understanding of the mechanisms of local immune responses in fish.