title: Corneal haze phenotype in Aldh3a1-null mice: In vivo confocal microscopy and tissue imaging mass spectrometry.
abstract: ALDH3A1 is a corneal crystallin that protects ocular tissues from ultraviolet radiation through catalytic and non-catalytic functions. In addition, ALDH3A1 plays a functional role in corneal epithelial homeostasis by simultaneously modulating proliferation and differentiation. We have previously shown that Aldh3a1 knockout mice in a C57B6/129sV mixed genetic background develop lens cataracts. In the current study, we evaluated the corneal phenotype of Aldh3a1 knockout mice bred into a C57B/6J congenic background (KO). In vivo confocal microscopy examination of KO and wild-type (WT) corneas revealed KO mice to exhibit corneal haze, manifesting marked light scattering from corneal stroma. This corneal phenotype was further characterized by Imaging Mass Spectrometry (IMS) with spatial resolution that revealed a trilayer structure based on differential lipid localization. In these preliminary studies, no differences were observed in lipid profiles from KO relative to WT mice; however, changes in protein profiles of acyl-CoA binding protein (m/z 9966) and histone H4.4 (m/z 11308) were found to be increased in the corneal epithelial layer of KO mice. This is the first study to use IMS to characterize endogenous proteins and lipids in corneal tissue and to molecularly explore the corneal haze phenotype. Taken together, the current study presents the first genetic animal model of cellular-induced corneal haze due to the loss of a corneal crystallin, and strongly supports the notion that ALDH3A1 is critical to cellular transparency. Finally, IMS represents a valuable new approach to reveal molecular changes underlying corneal disease.
authors.affiliation: Department of Environmental Health Sciences, Yale School of Public Health, New Haven, CT, 06521, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Ophthalmology, University of California-Irvine, Irvine, CA, 92697, USA.
authors.forename: James V
authors.initials: JV
authors.lastname: Jester
authors.affiliation: Department of Biochemistry and Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Anderson
authors.affiliation: U.S. Environmental Protection Agency, National Exposure Research Laboratory, 960 College Station Road, Athens, GA 30605, USA.
authors.forename: Satori A
authors.initials: SA
authors.lastname: Marchitti
authors.affiliation: Department of Biochemistry and Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, TN, 37232, USA.
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Schey
authors.affiliation: Department of Clinical Pharmacy, University of Colorado Anschutz Medical Campus, Aurora, CO, 80045, USA.
authors.forename: David C
authors.initials: DC
authors.lastname: Thompson
authors.affiliation: Department of Environmental Health Sciences, Yale School of Public Health, New Haven, CT, 06521, USA. Electronic address: vasilis.vasiliou@yale.edu.
authors.forename: Vasilis
authors.initials: V
authors.lastname: Vasiliou
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000172:deficiency / Q000235:genetics*; D000818:Animals; D003315:Cornea / Q000737:chemistry / Q000378:metabolism* / Q000473:pathology; D003316:Corneal Diseases / Q000378:metabolism / Q000473:pathology; D003319:Corneal Stroma / Q000502:physiology; D026261:Diazepam Binding Inhibitor / Q000737:chemistry / Q000378:metabolism; D004195:Disease Models, Animal; D000067493:Dynamic Light Scattering; D004848:Epithelium / Q000502:physiology; D019573:Epithelium, Corneal / Q000502:physiology; D006657:Histones / Q000737:chemistry / Q000378:metabolism; D007908:Lens, Crystalline / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D010641:Phenotype; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000444:Aldehyde Dehydrogenase; D026261:Diazepam Binding Inhibitor; D006657:Histones; D008055:Lipids; C105543:Aldh3a1 protein, mouse
keywords: ALDH3A1; Cellular transparency; Confocal microscope; Cornea haze; Imaging mass spectrometry