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Factors influencing the retrograde labeling of retinal ganglion cells with Fluorogold in animal optic nerve crush models |
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| Factors influencing the retrograde labeling of retinal ganglion cells with Fluorogold in animal optic nerve crush models |
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作者：Tzu-Lun … 文章来源：Department of Ophthalmology, Buddhist Tzu Chi General Hospital, Taiwan 点击数：281 更新时间：2011/9/13 
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PURPOSE: Fluorogold (FG) retrograde labeling of retinal ganglion cells (RGCs) is well-used in optic nerve (ON) crush. We investigated the impact factors influencing the RGC survival rate after ON crush.
METHODS: We standardized the ON crush in rats with a microvascular clip and comparing the crush effect with different crush methods. We also compared the RGCs counting between different FG injecting times, which was before sacrifice of animals (Group A) or before crush (Group B). Double stain with CD11b and FG was conducted in retinal sections.
RESULTS: Setting the crush time at 30 sec did not influence the flow of FG from the proximal part to the distal part of crush site [58.64±12.94/ high power field (HPF) and 67.86 ±12.11/HPF, respectively (p >0.05)]. Two weeks after a standardized ON crush, survival rate of RGCs decreased to 36.4% in the central retina and 27.7% in the mid-peripheral retina (Group A); survival rate of RGCs decreased to 59.2% in the central retina and 56.2% in the mid-peripheral retina (Group B). In double staining, FG and CD11b merged staining cells in Group B was higher than that of group A (34± 11% and 4± 3%, respectively, p<0.05). Over-counting of RGC density in Group B was resulted due to additional FG positive microglia.
CONCLUSIONS: Each lab should test their standard crush model to avoid factors influencing the FG labeling efficiency. FG injection one week before animal sacrifice is much close to the real RGC survival after ON crush than injections before ON crush. |
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