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Editors Selection IGR 8-3
Basic Research - RGC: RGC markers in optic neuropathy models
Comment by Francesca Cordeiro on:
48698 Quantitative analysis of retinal ganglion cell survival with Rbpms immunolabeling in animal models of optic neuropathies, Kwong JM; Quan A; Kyung H et al., Investigative Ophthalmology and Visual Science, 2011; 52: 9694-9702
Rbpms has been recently characterized by the authors (Kwong, Piri and Caprioli) as a new marker for RGCs, with better specificity than existing immune-markers including Brn-3. Although retrograde labeling is still regarded as the gold standard methodology for RGC identification, it does not adequately differentiate between RGC soma/ axonal loss and transport deficiencies. Likewise, there are problems with immunostaining in injured retinas, with only a small proportion of Brn3a labeling being seen in such eyes.
The paper describes validation of Rbpms in different glaucoma-models including excitoxicity, axotomy and laser-induced OHT, compared to labeling with retrograde Fluorogold (FG), and beta-III Tubulin as determined by histological analysis.
The paper shows some impressive colocalization of Rbpms with beta-III Tubulin on cross sections, although there is no statistical analysis of this in the whole retina mounts which would have been very much more convincing. Likewise, a merged figure of FG and Rbpms would have been nice to see, to allow the matching of cellular stains. However, this does not detract from the convincing data provided regarding the temporal and dose-dependent loss of RGCs seen in both the NMDA and ONT models. In addition, the correlation of IOP with RGC loss is very interesting, and agrees with previous studies on this subject. An added dimension in the analysis is the pattern with dark and light phases. This may be something that other groups may need to also consider in similar studies of OHT models. The greatest advantage of Rbpms appears to be its robustness in injury, and its ability to identify RGCs with intact axonal transport. It would be important for this to be investigated by other research groups in the area, to validate Rbpms as a specific marker in different models of RGC disease. In addition, it would be important to compare anterograde labeling techniques.
In summary, Rbpms provides an exciting new tool for assessing RGC counts in histological analysis, with further work needed to confirm its potential.
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