This research program continues to explore, develop and exploit the information content of Raman/resonance Raman/fluorescence spectra of human and animal lenses. The major thrust of this proposal is the application of our new instrumentation and techniques (automated two-dimensional scanning microprobe system) to study normal and cataractous lenses.
The specific aims are: 1) To obtain the precise distribution profiles of all the identifiable fluorophors in human lenses (normal, diabetic, Tibet/Beijing cataracts etc.) with continuously tunable laser excitation wavelenghts (300-800 nm); 3) To obtain the precise distribution profiles of important lens constituents such as -SH, S- S, Trp, H20 and Tyr which have been implicated in cataractogenesis; 3) To establish if certain fluorophors in the human lens are generated by photocatalyzed reactions; 4) To investigate the similarities and difference in fluorophor production between human and animal species; 5) To investigate the time-dependent spectral changes of glutathione-depleted intact lens (treated with 1-chloro-2,4-dinitrobenzene) to oxidative stress (H2O2 and superoxide anions) by multichannel """"""""snap-shot"""""""" technique; 6) To apply the precision difference Raman technqiue to detect subtle structural changes of lens crystallins upon oxidation and in the presence of calcium ion; 7) To investigate the in vitro behavior of the protein -SH groups in rat lens crystallins from very young rat lenses (17-day old); 8) To compare the reactivities of protein -SH groups with glutathione, cysteines, 2- mercaptoethanol and p-hydroxymercuribenzoate between bovine and rat crystallins; 9) To investigate if the following systems capable of producing red fluorophors: (1) Schiffbase adducts from cross-linking of carbonyl groups of malondialdehyde with the reactive amino groups of phospholipids and (ii) browning products from nonenzymatic glycosylation of lens crystallins; 10) To improve the sensitivity and the speed of data collection/transfer (approximately 10 times) in our automated scanning microprobe system.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY001746-14
Application #
3256178
Study Section
Visual Sciences A Study Section (VISA)
Project Start
1979-05-01
Project End
1993-04-30
Budget Start
1989-05-01
Budget End
1990-04-30
Support Year
14
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Georgia Institute of Technology
Department
Type
Schools of Arts and Sciences
DUNS #
097394084
City
Atlanta
State
GA
Country
United States
Zip Code
30332
Lo, W K; Kuck, J F; Shaw, A P et al. (1993) The altricial pigeon is born blind with a transient glycogen cataract. Exp Eye Res 56:121-6
Castillo, C G; Lo, W K; Kuck, J F et al. (1992) Nature and localization of avian lens glycogen by electron microscopy and Raman spectroscopy. Biophys J 61:839-44
Yu, N T; Cai, M Z; Lee, B S et al. (1991) Resonance Raman detection of a carotenoid in the lens of the deep-sea hatchetfish. Exp Eye Res 52:475-9
Zigman, S; Paxhia, T; McDaniel, T et al. (1991) Effect of chronic near-ultraviolet radiation on the gray squirrel lens in vivo. Invest Ophthalmol Vis Sci 32:1723-32
Chen, W L; Nie, S M; Kuck Jr, J F et al. (1991) Near-infrared Fourier transform Raman and conventional Raman studies of calf gamma-crystallins in the lyophilized state and in solution. Biophys J 60:447-55
Kuck, J F (1990) Late onset hereditary cataract of the emory mouse. A model for human senile cataract. Exp Eye Res 50:659-64
Yu, N T; Bando, M; Kuck Jr, J F (1990) Localization of UV-induced changes in mouse lens. Exp Eye Res 50:327-9
Nie, S M; Bergbauer, K L; Kuck Jr, J F et al. (1990) Near-infrared Fourier transform Raman spectroscopy in human lens research. Exp Eye Res 51:619-23
Cai, M Z; Kuck Jr, J F; Yu, N T (1989) Galactose-induced cataract in rat: Raman detection of sulfhydryl decrease and water increase along an equatorial diameter. Exp Eye Res 49:531-41
Yu, N T; Barron, B C; Kuck Jr, J F (1989) Distribution of two metabolically related fluorophors in human lens measured by laser microprobe. Exp Eye Res 49:189-94

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