Here, PFA immersion fixation followed by tissue processing and cryostat sectioning worked very well for the 14 antibodies investigated. bFGF (Mller cells and displaced amacrine CX-6258 HCl cells), GFAP (Mller cells and astrocytes), and vimentin (Mller 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 CX-6258 HCl 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 offered here will be applicable in fixed, cryosectioned pig retina. (J Histochem Cytochem 58:377C389, 2010) strong class=”kwd-title” Keywords: pig retina, photoreceptors, rods, cones, horizontal cells, bipolar cells, amacrine cells, ganglion cells, CX-6258 HCl retinal pigment epithelium, Mller cells The pig is an attractive non-primate model for exploring preclinical efficacy and conducting security trials for novel surgical and pharmaceutical therapies for various human diseases, as it is usually phylogenetically close to the human and shares many aspects of gross anatomy. Also, the pig vision and retina share many similarities with those of the human and are even more comparable than those of other large non-primate mammals (Beauchemin 1974; Ruiz-Ederra et al. 2005). Since the pig vision and retina closely resemble their human counterparts in size, number and distribution of rods and cones, shape, vasculature, and function, surgical and diagnostic gear used in the medical center can readily be applied (Ghosh and Arnr 2002; Hendrickson and Hicks 2002; Kiilgaard et al. 2002; Mahmoud et al. 2003; Ruiz-Ederra et al. 2005; Warfvinge et al. 2005; Iandiev et al. 2006; Lalonde et al. 2006; Lassota et al. 2006). The application of human ophthalmological diagnostics to the pig, such CX-6258 HCl as optical coherence tomography, corneal topography imaging, and multifocal electroretinogram, allow for behavioral measurements (Janknecht et al. 2001; Lalonde et al. 2006; Voss-Kyhn et al. 2007; Kyhn et al. 2008,2009a,b; Ng et al. 2008). Today a range of in vitro and in vivo porcine models with pathophysiological hallmarks of human retinal disorders are available (Chader 2002). For example, glaucoma models characterized by a loss of ganglion cells have been established, using acute (Kyhn et al. 2009b) or chronic elevated intraocular pressure (Ruiz-Ederra et al. 2005). A transgenic model of the inherited disease retinitis pigmentosa, characterized by a loss of photoreceptors, has been established (Petters et al. 1997; Li et al. 1998). Also, a transgenic pig with expression of green fluorescent protein has been produced and utilized for isolation of retinal stem cells, which have subsequently been used in transplantation studies (Park et al. 2001; Klassen et al. 2008). Retinal conditions such as retinal detachment (Scholda et al. 1999) and proliferative vitreoretinopathy (Garca-Layana et al. 1997) can also be modeled. The possibility of making organotypic cultures of embryonic and adult pig retina offers an experimental tool for many methods for retinal research (Gaudin et al. 1996; Luo et al. 2001; Garca et al. 2002; Winkler et al. 2002; Engelsberg et al. 2005; Kaempf et al. 2008). Studies of retinal development as well as evaluation of neuropathological changes in the diseased retina are preferably based on histological and immunohistochemical methods. For the pig retina, some cell typeCspecific antibodies have already been used in retinogenesis, descriptive, and neuropathological studies (Luo et al. 2001; Garca et al. 2002; Ghosh and Arnr 2002; Hendrickson and Hicks 2002; Yang et al. 2002; Ghosh et al. 2004,2007; Ruiz-Ederra et al. 2004; Warfvinge et al. 2005,2006; Lee et al. 2006b; Klassen et al. 2007,2008; Ahn et al. 2009; Guduric-Fuchs et al. 2009). In Rabbit polyclonal to CIDEB most of the CX-6258 HCl previous reports, the focus is usually on one or a few retinal cell types and often one developmental stage. However, in the very recent statement by Guduric-Fuchs et al. (2009), a range of markers for different developmental stages of pig retina was explored. The aim of the present study was to contribute to present knowledge by extending the range of markers specific for neuronal and glial cells in the adult normal pig retina. Danish domestic mixed breed (Danish Landrace/Duroc/Hampshire/Yorkshire) pigs were utilized for evaluating the specificity of different mono- and polyclonal antibodies, using fixed and cryosectioned tissue. The following antibodies were evaluated for neuronal and glial cells, respectively: recoverin (cones and rods), Rho4D2 (rods), transducin- (cones), calbindin (horizontal cells), protein kinase C- (PKC-) (bipolar cells), parvalbumin (amacrine cells), NeuN (ganglion cells, displaced amacrines), basic fibroblast growth factor (bFGF) (Mller cells, amacrine, and displaced.
