title: Anterior segment alterations and comparative aqueous humor proteomics in the buphthalmic rabbit (an American Ophthalmological Society thesis).
abstract: OBJECTIVE
  To use an integrated proteohistologic approach to gain insight into the anterior segment alterations in the buphthalmic rabbit.
  
  
  METHODS
  Eyes from 2- and 5-year-old buphthalmic and normal rabbits (n=20) were studied histologically. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) of aqueous humor (AH) was used to determine differential protein expression between animal groups. Western blot and immunohistochemistry were performed on selected differentially expressed proteins identified by LC-MS/MS.
  
  
  RESULTS
  The buphthalmic rabbits manifested a mild clinical phenotype with typical angle anomalies that appeared progressive by histology. Significantly thickened Descemet's membrane (DM) and anterior lens capsule in all buphthalmic rabbits showed increased fibronectin and collagen-IV immunolabeling. LC-MS/MS applying stringent filtering criteria revealed significant differential expression of several AH proteins in these rabbits. The protein of interest in the 2-year-old group was histidine-rich glycoprotein, and those in the 5-year-old group included alpha-2-HS-glycoprotein, clusterin, apolipoprotein E, interphotoreceptor retinoid-binding protein, transthyretin, cochlin, gelsolin, haptoglobin, hemopexin, and beta-2 microglobulin. The proteomic data for selected proteins was validated by Western blot and immunohistochemistry. A wide range of functional groups were affected by the altered AH proteins. These included extracellular matrix modulation, regulation of apoptosis, oxidative stress, and protein transport.
  
  
  CONCLUSIONS
  Multiple anterior segment alterations were histologically identified in the buphthalmic rabbits that showed progressive changes with age. The differentially expressed AH proteins in these rabbits suggest a multifunctional role for AH in modulating pathologic changes in DM, anterior lens capsule, and the angular meshwork in these animals.
authors.affiliation: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Deepak P
authors.initials: DP
authors.lastname: Edward
authors.forename: Rachida
authors.initials: R
authors.lastname: Bouhenni
mesh_terms: D000818:Animals; D000869:Anterior Eye Segment / Q000378:metabolism* / Q000473:pathology; D001082:Aqueous Humor / Q000737:chemistry / Q000378:metabolism*; D015153:Blotting, Western; D016022:Case-Control Studies; D002853:Chromatography, Liquid; D004195:Disease Models, Animal; D005136:Eye Proteins / Q000032:analysis / Q000378:metabolism*; D006871:Hydrophthalmos / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D007429:Intraocular Pressure / Q000502:physiology; D020543:Proteome / Q000032:analysis / Q000378:metabolism*; D040901:Proteomics; D011817:Rabbits; D053719:Tandem Mass Spectrometry
chemical_list: D005136:Eye Proteins; D020543:Proteome