Identification and localization of major soluble vitreous proteins in human ocular tissue.
OBJECTIVE
To identify the major soluble proteins from human vitreous, and to establish a baseline for comparison of vitreous samples from eyes with various diseases.


METHODS
Laboratory investigation.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Wu
Charlie W
CW
Department of Ophthalmology, University of Minnesota, Minneapolis, Minnesota 55455, USA.
Sauter
Jennifer L
JL
Johnson
Patrick K
PK
Chen
Ci-Di
CD
Olsen
Timothy W
TW
D000595:Amino Acid Sequence; D015153:Blotting, Western; D003930:Diabetic Retinopathy / Q000378:metabolism* / Q000601:surgery; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D008969:Molecular Sequence Data; D012160:Retina / Q000378:metabolism*; D012995:Solubility; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014019:Tissue Donors; D014821:Vitrectomy; D014822:Vitreous Body / Q000378:metabolism*
D005136:Eye Proteins