Glycosaminoglycans are part of amyloid fibrils: Ultrastructural evidence in avian AA amyloid stained with cuprolinic blue and labeled with immunogold

In domestic brown layer fowl, reactive amyloidosis of internal organs, such as liver and spleen, and of the joints is a common disorder. In a variety of amyloid types including the AA-amyloid of the chicken, in addition to amyloid fibrils, proteoglycans and glycosaminoglycans (GAGs) are found on immunohistochemistry or after extraction. The aim of the present report is to study amyloid fibrils for the ultrastructural location of GAGs by cuprolinic blue staining and immunogold labeling. Rabbit antichicken AA antiserum was used for the immunogold labeling on conventionally embedded and cryoembedded liver tissue and revealed similar results. Therefore conventional blocks could be used for further analysis. Cuprolinic blue staining was performed on blocks of joint tissue in which clearly discernable rod-shaped glycoproteins were encountered in between collagen fibrils. Moreover, it appeared to stain larger deposits which might represent amyloid. Postlabeling with the immunogold method of the cuprolinic blue-stained tissue proved that cuprolinic blue positive fibrils represented AA-amyloid fibrils. Therefore, it was concluded that the GAGs which appeared to colocalize with the fibrillar microanatomy of amyloid, represent a structural part of the amyloid fibrils and that the avian amyloid fibrils may be considered as a pathological proteoglycan.