Quantitative analysis of retinal ganglion cell (RGC) loss in aging DBA/2NNia glaucomatous mice: comparison with RGC loss in aging C57/BL6 mice.
OBJECTIVE
To quantify the extent and pattern of retinal ganglion cell (RGC) loss in the DBA2/NNia glaucomatous mouse strain as a function of age and compare it with ganglion cell loss in a nonglaucomatous strain.


METHODS
All the ganglion cells in retinas of DBA/2NNia and C57/BL6 mice of various ages (five eyes per age group in 3-month intervals from 3 to 18 months of age) were counted. A novel counting method that does not rely on sampling and that uses retrograde labeling of RGCs with Fluorogold (Fluorochrome; Englewood, CO) was used. RGC loss in the glaucomatous DBA/2NNia mouse strain was contrasted to RGC loss in C57 mice at the same ages. The total number of Fluorogold-labeled cells per retina was compared within and among the two strains as a function of age. In addition, RGC density maps were constructed for each retina, and the range of densities for each age group was compared within and among the two strains. IOP in awake, nonsedated DBA/2NNia mice was measured with a rebound tonometer.


RESULTS
RGC loss started between 12 and 15 months of age in C57 mice and led to an approximate 46% reduction by 18 months of age. The rate of loss was best approximated by a second-order polynomial curve. In comparison, DBA/2NNia mice also began showing RGC loss at approximately 12 months of age, but it proceeded at a much faster rate, with approximately 64% of their RGCs dying by the 15th month of age but little additional loss thereafter. RGC loss in the DBA animals had a focal pattern that appeared more patchy and showed greater variability than the age-related loss in C57 mice, which was more diffuse. IOP and total retinal area in DBA/2NNia mice began to increase at approximately 6 months of age. IOP normalized after the 12th month of age.


CONCLUSIONS
Age-related RGC loss of up to 50% can occur in the C57 mouse by 18 months of age. The loss does not proceed linearly with age, as is often assumed, and differs both in extent and locational pattern from pathologic RGC loss secondary to glaucoma in DBA/2NNia mouse retinas.
Danias
John
J
Department of Ophthalmology, Mount Sinai School of Medicine, Mount Sinai Medical Center, 1 Gustave L. Levy Place, New York, NY 10029, USA. john.danias@mssm.edu
Lee
Kevin C
KC
Zamora
Maria-Florencia
MF
Chen
Bin
B
Shen
Fran
F
Filippopoulos
Theodoros
T
Su
Yanling
Y
Goldblum
David
D
Podos
Steven M
SM
Mittag
Thom
T
D000375:Aging / Q000502:physiology*; D000818:Animals; D002452:Cell Count; D016923:Cell Death; D005456:Fluorescent Dyes; D005901:Glaucoma / Q000150:complications*; D007429:Intraocular Pressure; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D009896:Optic Atrophy / Q000209:etiology; D012165:Retinal Ganglion Cells / Q000473:pathology*; D013266:Stilbamidines
C049774:2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt; D005456:Fluorescent Dyes; D013266:Stilbamidines