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Y. Takihara, M. Inatani, K. Iwao, M. Iwao, H. Tanihara; Live Imaging of Axonal Transport in Retinal Ganglion Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):5470.
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© ARVO (1962-2015); The Authors (2016-present)
Elevated intraocular pressure causes backbowing of lamina cribrosa. Previous histological studies revealed that this backbowing results in obstruction of retrograde axonal transport in retinal ganglion cells (RGCs). However, no studies have shown live imaging of axonal transport in RGCs.
RGCs from retina of 6 to 8 day-old DA rats were purified by a two-step immunopanning procedure. After 11 days, to examine whether synaptogenesis is induced, immunocytochemical staining with an anti-synaptophysin antibody was carried out. Then, cDNA construct for mouse brain-derived neurotrophic factor (BDNF) tagged with GFP was transfected to RGCs with lipofectamine 2000. We observed live imaging of the movement of GFP-tagged BDNF in the axon with time-lapse recording. To distinguish an axon from dendrites, immunocytochemistry was performed with an anti-MAP2 antibody.
Synaptophysin-positive neurites were observed in long-term cultivated RGCs. The time-lapse recording showed the anterograde and retrograde movements of punctate granules of transfected BDNF in the MAP2-negative axon.
Synaptogenesis is formed between the axons and dendrites from long-term cultivated RGCs. Time-lapse recording is useful for live imaging of axonal transport in RGCs.
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