Abstract
Purpose :
Evaluate presumptive amyloidosis by Congo red staining under fluorescent microscopy and identify the presence of β-amyloid by immunohistochemistry and western blot in cultured adult porcine retina.
Methods :
Conventional Congo red staining under red fluorescence was used to determine the presence of amyloid proteins in experimentally-induced injuries in cultured retina. Immunohistochemical labeling and western blot were used to investigate the presence of β-amyloid. Co-cultures, either of retina with human retinal pigment epithelial (RPE) cells or retina with human neural progenitors, were performed to investigate if amyloidosis could be reduced.
Results :
Congo red staining appeared as fluorescent puncta in the adult porcine retina after three days of culture. Congo red puncta were particularly associated with the outer nuclear layer, often adjacent to tissue cavities. The occurance of Congo red puncta appeared to be reduced in co-cultured specimens. Immunohistochemistry and western blot indicated the presence of amyloid protein precursors as well as cleaved β-amyloid.
Conclusions :
Our data collectively suggest that amyloidosis might be included in the retinal degeneration that is induced during culture. It is also suggested that β-amyloid formation might participate in this process, and that trophic and structural support to the injured retina can reduce amyloidosis.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.