Time-Lapse Retinal Ganglion Cell Dendritic Field Degeneration Imaged in Organotypic Retinal Explant Culture

Investigative Ophthalmology & Visual Science
Thomas V JohnsonHarry A Quigley

Abstract

To develop an ex vivo organotypic retinal explant culture system suitable for multiple time-point imaging of retinal ganglion cell (RGC) dendritic arbors over a period of 1 week, and capable of detecting dendrite neuroprotection conferred by experimental treatments. Thy1-YFP mouse retinas were explanted and maintained in organotypic culture. Retinal ganglion cell dendritic arbors were imaged repeatedly using confocal laser scanning microscopy. Maximal projection z-stacks were traced by two masked investigators and dendritic fields were analyzed for characteristics including branch number, size, and complexity. One group of explants was treated with brain derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) added to the culture media. Changes in individual dendritic fields over time were detected using pair-wise comparison testing. Retinal ganglion cells in mouse retinal explant culture began to degenerate after 3 days with 52.4% surviving at 7 days. Dendritic field parameters showed minimal change over 8 hours in culture. Intra- and interobserver measurements of dendrite characteristics were strongly correlated (Spearman rank correlations consistently > 0.80). Statistically significant (P < 0.001) dendritic...Continue Reading

Citations

Jun 11, 2016·The European Journal of Neuroscience·Kate E BinleyJames E Morgan
Apr 4, 2019·Translational Vision Science & Technology·Rania A MasriUlrike Grünert
Mar 14, 2020·International Journal of Molecular Sciences·Ines RosignolPatricia Boya

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Methods Mentioned

BETA
confocal microscopy
transgenic
imaging techniques

Software Mentioned

Fiji
NeuronJ
ImageJ
Zen

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