The movement and morphological changes of single cells in the corneal stroma of the living mouse will be observed under a fluorescence microscope. The behavior of normal cells as well as those in the neighborhood of a mild, circumscribed, injury will be followed. The injury will consist of a small circular epithelial debridement or the introduction of hydrogen ions by iontophoresis over a 0.4 mm circle, both of which are reproducible. The keratocytes and other stromal cells will be followed by the use of transgenic mice engineered to express fluorescent proteins ubiquitously, or by staining with nuclear or cytoplasmic fluorescent dyes. Leucocytes will be labeled with nuclear dye systemically. The images are recorded by a digital camera and time-lapse sequences are constructed for analysis. The mechanisms of cell death, activation and repopulation after a mild injury, will be characterized and analyzed. It is intended that further studies will lead to a clarification of the entire wound healing process in the cornea. It has an immediate relevance to the healing of surgical incisions and laser ablations made in the course of refractive surgery. Since the reactions of cells to injury in all tissues is likely to be similar, the results should have a wide significance.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY000431-32
Application #
6603732
Study Section
Visual Sciences A Study Section (VISA)
Program Officer
Fisher, Richard S
Project Start
1994-02-01
Project End
2004-12-31
Budget Start
2003-07-01
Budget End
2004-12-31
Support Year
32
Fiscal Year
2003
Total Cost
$204,375
Indirect Cost
Name
Columbia University (N.Y.)
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
Nagasaki, Takayuki; Zhao, Jin (2005) Uniform distribution of epithelial stem cells in the bulbar conjunctiva. Invest Ophthalmol Vis Sci 46:126-32
Smith, R T; Chan, J K; Nagasaki, T et al. (2005) A method of drusen measurement based on reconstruction of fundus background reflectance. Br J Ophthalmol 89:87-91
Fischbarg, Jorge; Maurice, David M (2004) An update on corneal hydration control. Exp Eye Res 78:537-41
Maurice, D M; Zhao, J; Nagasaki, T (2004) A novel microscope system for time-lapse observation of corneal cells in a living mouse. Exp Eye Res 78:591-7
Zhao, Jin; Nagasaki, Takayuki (2004) Mechanical damage to corneal stromal cells by epithelial scraping. Cornea 23:497-502
Zhao, Jin; Nagasaki, Takayuki (2003) Lacrimal gland as the major source of mouse tear factors that are cytotoxic to corneal keratocytes. Exp Eye Res 77:297-304
Nagasaki, Takayuki; Zhao, Jin (2003) Centripetal movement of corneal epithelial cells in the normal adult mouse. Invest Ophthalmol Vis Sci 44:558-66
Maurice, David M (2002) Drug delivery to the posterior segment from drops. Surv Ophthalmol 47 Suppl 1:S41-52
Maurice, D (2001) Review: practical issues in intravitreal drug delivery. J Ocul Pharmacol Ther 17:393-401
Zhao, J; Nagasaki, T; Maurice, D M (2001) Role of tears in keratocyte loss after epithelial removal in mouse cornea. Invest Ophthalmol Vis Sci 42:1743-9

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