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

Abstract

S U M M A R Y 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-g (cones), ROM-1 (photoreceptor outer segments), calbindin (horizontal cells), PKC-a (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)

N-methyl-N-nitrosourea-induced neuronal cell death in a large animal model of retinal degeneration in vitro

“…Retinas from the two in vivo control eyes displayed normal morphology as previously described (Taylor et al, 2013a;Li et al, 1998;Engelsberg et al, 2005;Johansson et al, 2010). Hematoxylin and eosin staining (HTX) showed distinct lamination and wellpopulated cell layers (Fig.…”

Proliferative Vitreoretinopathy in the Swine–A New Model

“…The swine eye has become an attractive non-primate model in experimental studies because of its close similarity to the human eye in size, in selective concentration of cone photoreceptors in the visual streak, and its holangiotic retinal vascular pattern. [20][21][22] In this paper, we present a new model of PVR using the swine. Green fluorescent protein-positive (GFPþ) RPE cells were injected intravitreally so that the fate of these cells could be traced during the development of PVR and their contribution to the formation of epiretinal membranes documented.…”

Novel Cellular Bouton Structure Activated by ATP in the Vascular Wall of Porcine Retinal Arterioles

“…These cell bodies may potentially be related to pericytes or may be of neuronal or glial origin, which is in accordance with recent findings of purinergic signaling in these cells in the retina. 7 The lack of immunoreactivity in the newly identified boutons may explain the lack of focus on these structures in the literature [22][23][24] and prompts further studies to identify the cells from which they originate.…”