Immunohistochemical and Special Staining Techniques for the Investigation of Inflammatory and Degenerative Changes in the Ear of the Harbor Porpoise (Phocoena phocoena)
To facilitate detailed investigation of inflammatory and degenerative changes of harbor porpoise (Phocoena phocoena) ears, immunohistochemical and special staining techniques were implemented. One polyclonal antibody and three monoclonal markers of different species specificities directed against different antigens were tested for immunohistochemical cross-reactivity on formalin-fixed, paraffin-embedded middle and inner ear sections of a harbor porpoise with Stenurus minor infection. Additionally, Luxol Fast Blue, Bielschofsky and Cresyl Violet special staining for the detection of degenerative changes of nerve fibers, neurons, and chromatolysis, respectively, were performed. All investigated antibodies showed specific immunoreactivity with leukocytes in harbor porpoise ears. Lymphocytes were labeled by a polyclonal human-specific anti-CD3 antibody. Cells of the histiocytic lineage were recognized by a monoclonal human-specific antibody. The MHC class II-antigen was labeled by a human-specific monoclonal antibody. A positive species cross-reactivity of the human-specific monoclonal anti-CD79α-marker was observed. Myelin and myelinated axons were stained by Luxol Fast Blue staining. Axons were further visualized by Bielschofsky special staining, and Nissl substance was detected in the cytoplasm of neurons by Cresyl Violet staining. This study shows the applicability of leukocyte markers for the characterization of inflammatory responses in formalin-fixed, paraffin embedded tissue sections of the ear of the harbor porpoise by immunohistochemistry. Additionally performed special staining techniques enable the detection of neuronal structures and degenerative changes in harbor porpoise ears.
This study is funded by the Federal Ministry of Food, Agriculture and Consumer Protection (Bundesministerium für Ernährung, Landwirtschaft und Verbraucherschutz), through the Federal Agency of Agriculture and Food (Bundesanstalt für Landwirtschaft und Ernährung).