title: Advanced Clinical Imaging and Tissue-based Biomarkers of the Eye for Toxicology Studies in Minipigs.
abstract: There is increased interest to use minipigs in ocular toxicology studies due to their anatomical similarities with human eyes and as a substitute for nonhuman primates. This requires adaptation of enhanced optical coherence tomography (OCT) techniques and of ocular relevant immunohistochemistry (IHC) or in situ hybridization (ISH) markers to porcine eyes. In this study, OCT and OCT angiography (AngioOCT) were performed on adult Göttingen minipigs. To increase structural information on retinal and choroidal vasculature, OCT data were speckle denoized and choroidal blood vessels were segmented with threshold filtering. In addition, we established a set of IHC and ISH markers on Davidson's fixed paraffin-embedded minipig eyes: neurofilament-160, neuronal nuclei, calretinin, protein kinase C-α, vimentin, glial fibrillary acidic protein, glutamine synthetase, ionized calcium-binding adaptor molecule-1, rhodopsin, synaptophysin, postsynaptic density protein-95, retinal pigment epithelium (RPE)-specific protein-65, von Willebrand factor, α-smooth muscle actin, desmin, and Ki-67, thus enabling visualization of retinal neuronal and glial cells, photoreceptors, synapses, RPE, blood vessels, myocytes, macrophages, or cell proliferation. Using ISH, transcripts of vascular endothelial growth factor A, angiopoietin-2, and endothelial tyrosine kinase were visualized. This article describes for the first time in minipig eyes speckle noise-free OCT, AngioOCT, and a set of IHC/ISH markers on Davidson's fixed paraffin-embedded tissues and helps to establish the minipig for ocular toxicology and pharmacology studies.
authors.affiliation: Roche Innovation Center Basel, Basel, Switzerland elke-astrid.atzpodien@roche.com.
authors.forename: Elke-Astrid
authors.initials: EA
authors.lastname: Atzpodien
authors.affiliation: Roche Innovation Center Basel, Basel, Switzerland.
authors.forename: Bjoern
authors.initials: B
authors.lastname: Jacobsen
authors.affiliation: Roche Innovation Center Basel, Basel, Switzerland.
authors.forename: Juergen
authors.initials: J
authors.lastname: Funk
authors.affiliation: Roche Innovation Center Basel, Basel, Switzerland.
authors.forename: Bernd
authors.initials: B
authors.lastname: Altmann
authors.affiliation: Roche Innovation Center Penzberg, Penzberg, Germany.
authors.forename: Manuel A
authors.initials: MA
authors.lastname: Silva Munoz
authors.affiliation: Roche Innovation Center Basel, Basel, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Singer
authors.affiliation: OCT Research Laboratory, Department of Ophthalmology, University of Basel, Basel, Switzerland.
authors.forename: Cyrill
authors.initials: C
authors.lastname: Gyger
authors.affiliation: OCT Research Laboratory, Department of Ophthalmology, University of Basel, Basel, Switzerland.
authors.forename: Pascal
authors.initials: P
authors.lastname: Hasler
authors.affiliation: OCT Research Laboratory, Department of Ophthalmology, University of Basel, Basel, Switzerland.
authors.forename: Peter
authors.initials: P
authors.lastname: Maloca
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis*; D005123:Eye / Q000737:chemistry* / Q000000981:diagnostic imaging*; D005260:Female; D007150:Immunohistochemistry; D008297:Male; D012160:Retina / Q000000981:diagnostic imaging; D012171:Retinal Vessels / Q000000981:diagnostic imaging; D013552:Swine; D013556:Swine, Miniature; D041623:Tomography, Optical Coherence
chemical_list: D015415:Biomarkers
keywords: Davidson’s fixative; OCT angiography; eye; immunohistochemistry; in situ hybridization; minipig; optical coherence tomography