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Uptake of Retrograde Tracers by Intact Optic Nerve Axons: A New Way to Label Retinal Ganglion Cells.

Liang YX, Yang J, Yuan TF, So KF - PLoS ONE (2015)

Bottom Line: Here we report that an intact optic nerve could uptake retrograde tracers applied onto the surface of the nerve, leading to high efficiency labelling of ganglion cells in the retina with long-term survival of cells.This finding provides an alternative way to label retinal ganglion cells without damaging the optic tract.This will facilitate anatomical studies in identifying the morphology and connectivity of retinal ganglion cells, allowing secondary or triple labelling manipulations for long-term investigations.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Brain and Cognitive Sciences, the University of Hong Kong, Hong Kong, China; Department of Anatomy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.

ABSTRACT
Retrograde labelling of retinal ganglion cells with optic nerve transection often leads to degeneration of ganglion cells in prolonged experiments. Here we report that an intact optic nerve could uptake retrograde tracers applied onto the surface of the nerve, leading to high efficiency labelling of ganglion cells in the retina with long-term survival of cells. This method labelled a similar number of ganglion cells (2289±174 at 2 days) as the retrograde labeling technique from the superior colliculus (2250±94) or optic nerve stump (2279±114) after transection. This finding provides an alternative way to label retinal ganglion cells without damaging the optic tract. This will facilitate anatomical studies in identifying the morphology and connectivity of retinal ganglion cells, allowing secondary or triple labelling manipulations for long-term investigations.

No MeSH data available.


Related in: MedlinePlus

Intact ON labelling approach results in the same quality of RGC filling with fluorescent dyes.2A: FG labelling of RGCs through superior colliculus application; 2B: FG labelling of RGCs through ON cut approach; 2C: FG labelling of RGCs through intact ON approach; 2D: GB labelling of RGCs through intact ON approach. Scale bar represents 20 μm.
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pone.0128718.g002: Intact ON labelling approach results in the same quality of RGC filling with fluorescent dyes.2A: FG labelling of RGCs through superior colliculus application; 2B: FG labelling of RGCs through ON cut approach; 2C: FG labelling of RGCs through intact ON approach; 2D: GB labelling of RGCs through intact ON approach. Scale bar represents 20 μm.

Mentions: We found that in our study, SC labelling and ON stump labelling are able to label most RGCs in the retina, as previously reported. Surprisingly, applying FG onto the surface of the ON was also sufficient for labelling of RGCs after 2 days of labelling (Fig 1), with the same levels of fluorescence intensity and soma filling of the RGCs (Fig 2). The intensity of fluorescence met the routine requirements for following cell counting and morphological studies. We also quantified the RGCs number per mm2 with different labelling methods (Table 1). At 2 days, the labeling efficacy of intact ON labeling was comparable to SC labeling and optic nerve stump labeling.


Uptake of Retrograde Tracers by Intact Optic Nerve Axons: A New Way to Label Retinal Ganglion Cells.

Liang YX, Yang J, Yuan TF, So KF - PLoS ONE (2015)

Intact ON labelling approach results in the same quality of RGC filling with fluorescent dyes.2A: FG labelling of RGCs through superior colliculus application; 2B: FG labelling of RGCs through ON cut approach; 2C: FG labelling of RGCs through intact ON approach; 2D: GB labelling of RGCs through intact ON approach. Scale bar represents 20 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4465746&req=5

pone.0128718.g002: Intact ON labelling approach results in the same quality of RGC filling with fluorescent dyes.2A: FG labelling of RGCs through superior colliculus application; 2B: FG labelling of RGCs through ON cut approach; 2C: FG labelling of RGCs through intact ON approach; 2D: GB labelling of RGCs through intact ON approach. Scale bar represents 20 μm.
Mentions: We found that in our study, SC labelling and ON stump labelling are able to label most RGCs in the retina, as previously reported. Surprisingly, applying FG onto the surface of the ON was also sufficient for labelling of RGCs after 2 days of labelling (Fig 1), with the same levels of fluorescence intensity and soma filling of the RGCs (Fig 2). The intensity of fluorescence met the routine requirements for following cell counting and morphological studies. We also quantified the RGCs number per mm2 with different labelling methods (Table 1). At 2 days, the labeling efficacy of intact ON labeling was comparable to SC labeling and optic nerve stump labeling.

Bottom Line: Here we report that an intact optic nerve could uptake retrograde tracers applied onto the surface of the nerve, leading to high efficiency labelling of ganglion cells in the retina with long-term survival of cells.This finding provides an alternative way to label retinal ganglion cells without damaging the optic tract.This will facilitate anatomical studies in identifying the morphology and connectivity of retinal ganglion cells, allowing secondary or triple labelling manipulations for long-term investigations.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Brain and Cognitive Sciences, the University of Hong Kong, Hong Kong, China; Department of Anatomy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.

ABSTRACT
Retrograde labelling of retinal ganglion cells with optic nerve transection often leads to degeneration of ganglion cells in prolonged experiments. Here we report that an intact optic nerve could uptake retrograde tracers applied onto the surface of the nerve, leading to high efficiency labelling of ganglion cells in the retina with long-term survival of cells. This method labelled a similar number of ganglion cells (2289±174 at 2 days) as the retrograde labeling technique from the superior colliculus (2250±94) or optic nerve stump (2279±114) after transection. This finding provides an alternative way to label retinal ganglion cells without damaging the optic tract. This will facilitate anatomical studies in identifying the morphology and connectivity of retinal ganglion cells, allowing secondary or triple labelling manipulations for long-term investigations.

No MeSH data available.


Related in: MedlinePlus