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John Danias, Fran Shen, David Goldblum, Bin Chen, Jerome Ramos-Esteban, Steven M. Podos, Thom Mittag; Cytoarchitecture of the Retinal Ganglion Cells in the Rat. Invest. Ophthalmol. Vis. Sci. 2002;43(3):587-594.
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purpose. To determine the number and cytoarchitecture of retinal ganglion cells
(RGCs) in the female Wistar rat, by using a newly devised procedure for
rapid RGC counting in the entire retina that avoids assumptions about
RGC spatial arrangement.
methods. RGCs of normal female Wistar rats were retrogradely labeled with a
fluorescent tracer. Automated counting was accomplished by
applying standard imaging software to analysis of all labeled cells in
retinal flatmounts. The method was validated by comparison of automated
and manual counts of 70,000 RGCs in frames covering the density range
in the normal rat retina of 600 to 3600 RGC/mm2. RGC
numbers were determined for each retina and compared with the
contralateral retina of the same animal. RGC density maps were
constructed for each retina. RGC size distribution was determined.
results. Automated RGC counting showed a good linear correlation with
manual counting (R 2 = 0.9416). Mean
total RGC count in 10 rat eyes was 97,609 ± 3,930 (SEM) per eye.
Contralateral eyes differed by an average of 4.1% (3983 ± 5098
RGCs). Size analysis calculated from cell areas confirmed that the
majority of rat RGCs are between 7 and 21.5 μm in equivalent
diameter. The RGC counts for all frames at the same eccentricity in all
10 of the retinas showed that variability increased with eccentricity
and increased further as the fractional area of the retina sampled at
each eccentricity was reduced. There was also significant variability
in the spatial density of the RGCs at the same eccentricity location
between different eyes. Comparison of total RGC counts between left and
right eyes estimated from RGC counts in sectors of the retina
(hemiretinas or quadrants) showed increased variability compared with
counting all the RGCs in a retina.
conclusions. RGCs in the Wistar rat display significant variability in their
cytoarchitecture. Such variability can make quantification by sampling
problematic for diffuse, and particularly, for focal RGC losses
resulting from experimental interventions, unless virtually the entire
RGC population is counted.
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