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A Battery of Cell- and Structure-specific Markers for the Adult Porcine Retina

Ulrica Englund Johansson, Sajedeh Eftekhari and Karin Warfvinge

Department of Clinical Sciences, Division of Ophthalmology, Lund University, Lund, Sweden

Correspondence to: Ulrica Englund Johansson, PhD, MSc, Department of Clinical Sciences, Division of Ophthalmology, Lund University BMC B13 22184, Lund, Sweden. E-mail: ulrica.englund_johansson{at}med.lu.se

The pig is becoming an increasingly used non-primate model in experimental studies of human retinal diseases and disorders. The anatomy, size, and vasculature of the porcine eye and retina closely resemble their human counterparts, which allows for application of standard instrumentation and diagnostics used in the clinic. Despite many reports that demonstrate immunohistochemistry as a useful method for exploring neuropathological changes in the mammalian central nervous system, including the pig, the porcine retina has been sparsely described. Hence, to facilitate further immunohistochemical analysis of the porcine retina, we report on the successful use of a battery of antibodies for staining of paraformaldehyde-fixed cryosectioned retina. The following antibodies were evaluated for neuronal cells and structures: recoverin (cones and rods), Rho4D2 (rods), transducin- (cones), ROM-1 (photoreceptor outer segments), calbindin (horizontal cells), PKC- (bipolar cells), parvalbumin (amacrine and displaced amacrine cells), and NeuN (ganglion cells and displaced amacrines). For detecting synaptic connections in fiber layers, we used an antibody against synaptobrevin. For detecting retinal pigment epithelium, we studied antibodies against cytokeratin and RPE65, respectively. The glial cell markers used were bFGF (Müller cells and displaced amacrine cells), GFAP (Müller cells and astrocytes), and vimentin (Müller cells). Each staining effect was evaluated with regard to its specificity, sensitivity, and reproducibility in the identification of individual cells, specific cell structures, and fiber layers, respectively. The markers parvalbumin and ROM-1 were tested here for the first time for the porcine retina. All antibodies tested resulted in specific staining of high quality. In conclusion, all immunohistochemical protocols presented here will be applicable in fixed, cryosectioned pig retina. (J Histochem Cytochem 58:377–389, 2010)

Key Words: pig retina • photoreceptors • rods • cones • horizontal cells • bipolar cells • amacrine cells • ganglion cells • retinal pigment epithelium • Müller cells

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