Two corneal diseases will be investigated: Macular corneal dystrophy: Abnormal glycoconjugates synthesized by corneas with macular dystrophy in organ culture will be characterized to establish the cause of the abnormal lectin binding pattern known to be present in corneas with macular dystrophy. In addition we will attempt to determine whether the defect in sulfation of keratan sulfate (KS) proteoglycan in corneas with macular dystrophy is due to the deficiency of KS-sulfotransferases in the diseased corneas. This study may enable us to elucidate the biochemical abnormalities responsible for the development of macular corneal dystrophy. Keratoconus: We have shown by 2D-electrophoresis that, compared to normal corneas, a number of additional proteins (K) are present and a number of normal corneal proteins (N) are absent in keratoconus corneas. The proposed study will attempt to determine whether the """"""""K"""""""" proteins in keratoconus have been derived from the """"""""N"""""""" proteins. Polyclonal antibodies will be prepared against one of the """"""""N"""""""" proteins. These antibodies will be used to: (1) stain the Western blots of 2D-gels of proteins from normal and keratoconus corneas to determine whether one or more of the """"""""K"""""""" proteins are related to the """"""""N"""""""" protein and (2) study the synthesis of """"""""N"""""""" and """"""""K"""""""" proteins by corneas in organ culture by immunoprecipitation to determine whether one or more """"""""K"""""""" proteins in keratoconus appears as a consequence of the abnormal processing of the normal corneal protein (N). The cDNA libraries of normal and keratoconus corneas will be prepared in lambda gtll expression vector and the nucleotide sequence of full-length cDNA inserts of the """"""""N"""""""" protein and its abnormal counterpart (K) in keratoconus, if found, will be determined. Amino acid sequence of the """"""""N"""""""" and """"""""K"""""""" proteins will be analyzed for homology with the sequences in the databases of the National Biomedical Research Foundation. We anticipate that this study will help us identify and characterize the proteins involved in the development of keratoconus. (See rebuttal to the previous summary statement, pages 20-22.)

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY004034-07A1
Application #
3258498
Study Section
Visual Sciences A Study Section (VISA)
Project Start
1981-06-01
Project End
1991-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
7
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Tufts University
Department
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02111
Panjwani, N; Zhao, Z; Baum, J et al. (1992) Acanthamoebae bind to glycolipids of rabbit corneal epithelium. Infect Immun 60:3460-3
Panjwani, N; Clark, B; Cohen, M et al. (1990) Differential binding of P. aeruginosa and S. aureus to corneal epithelium in culture. Invest Ophthalmol Vis Sci 31:696-701
Panjwani, N; Zaidi, T S; Gigstad, J E et al. (1990) Binding of Pseudomonas aeruginosa to neutral glycosphingolipids of rabbit corneal epithelium. Infect Immun 58:114-8
Panjwani, N; Michalopoulos, G; Song, J et al. (1990) Neutral glycolipids of migrating and nonmigrating rabbit corneal epithelium in organ and cell culture. Invest Ophthalmol Vis Sci 31:689-95
Panjwani, N; Drysdale, J; Clark, B et al. (1989) Protein-related abnormalities in keratoconus. Invest Ophthalmol Vis Sci 30:2481-7
Funderburgh, J L; Panjwani, N; Conrad, G W et al. (1989) Altered keratan sulfate epitopes in keratoconus. Invest Ophthalmol Vis Sci 30:2278-81
Panjwani, N; Baum, J (1988) A histochemical comparison of human corneal stromal glycoconjugates with eight other species. Distinct species-dictated differences in binding sites of Griffonia simplicifolia I. Histochemistry 89:41-5
Yue, B Y; Panjwani, N; Sugar, J et al. (1988) Glycoconjugate abnormalities in cultured keratoconus stromal cells. Arch Ophthalmol 106:1709-12
Panjwani, N; Rodrigues, M; Free, K et al. (1987) Lectin receptors of amyloid in corneas with lattice dystrophy. Arch Ophthalmol 105:688-91
Hsieh, P; Baum, J (1985) Effects of fibroblastic and endothelial extracellular matrices on corneal endothelial cells. Invest Ophthalmol Vis Sci 26:457-63