Identification and localization of major soluble vitreous proteins in human ocular tissue

To identify the major soluble proteins from human vitreous, and to establish a baseline for comparison of vitreous samples from eyes with various diseases. Laboratory investigation. Normal vitreous was obtained from eight human donor eyes and from eight eyes of patients undergoing diabetic vitrectom...

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Bibliographic Details
Published in:American journal of ophthalmology 2004-04, Vol.137 (4), p.655-661
Main Authors: Wu, Charlie W., Sauter, Jennifer L., Johnson, Patrick K., Chen, Ci-Di, Olsen, Timothy W.
Format: Article
Language:English
Subjects:
Acids
Amino Acid Sequence
Biological and medical sciences
Blotting, Western
Diabetes
Diabetic retinopathy
Diabetic Retinopathy - metabolism
Diabetic Retinopathy - surgery
Disease
Electrophoresis, Polyacrylamide Gel
Eye and associated structures. Visual pathways and centers. Vision
Eye Proteins - metabolism
Fluorescent Antibody Technique, Indirect
Fundamental and applied biological sciences. Psychology
Humans
Macular degeneration
Mass spectrometry
Molecular Sequence Data
Molecular weight
Proteins
Retina - metabolism
Sodium
Solubility
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Tissue Donors
Vascular endothelial growth factor
Vertebrates: nervous system and sense organs
Vitrectomy
Vitreous Body - metabolism
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Summary:To identify the major soluble proteins from human vitreous, and to establish a baseline for comparison of vitreous samples from eyes with various diseases. Laboratory investigation. Normal vitreous was obtained from eight human donor eyes and from eight eyes of patients undergoing diabetic vitrectomy. Vitreous specimens were subjected to SDS-PAGE and MALDI-TOF-MS analysis. Six specific antibodies were used to identify proteins using Western blot. Four proteins were localized within ocular tissue in a normal donor eyebank eye. We found eight distinct bands on SDS-PAGE in normal vitreous and two additional bands (hemoglobin) in eyes with diabetic retinopathy. Proteins were identified using MALDI-TOF-MS and confirmed by Western blot. We established a quantitative analysis of relative protein concentrations of undiluted vitreous. Immunohistochemistry localized selected proteins within the posterior segment layers. We present a normal human vitreous protein profile using current technologies and provide a baseline for comparison to ocular disease states. Tissue distribution of vitreous proteins may help to elucidate more specific protein function.
ISSN:0002-9394
1879-1891
DOI:10.1016/j.ajo.2003.11.009