May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Live Imaging of Axonal Transport in Retinal Ganglion Cells
Author Affiliations & Notes
  • Y. Takihara
    Dept of Ophthal & Vis Science, Kumamoto Univ Sch of Med, Kumamoto, Japan
  • M. Inatani
    Dept of Ophthal & Vis Science, Kumamoto Univ Sch of Med, Kumamoto, Japan
  • K. Iwao
    Dept of Ophthal & Vis Science, Kumamoto Univ Sch of Med, Kumamoto, Japan
  • M. Iwao
    Dept of Ophthal & Vis Science, Kumamoto Univ Sch of Med, Kumamoto, Japan
  • H. Tanihara
    Dept of Ophthal & Vis Science, Kumamoto Univ Sch of Med, Kumamoto, Japan
  • Footnotes
    Commercial Relationships  Y. Takihara, None; M. Inatani, None; K. Iwao, None; M. Iwao, None; H. Tanihara, None.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 5470. doi:https://doi.org/
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    • Get Citation

      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. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : 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.

Methods: : 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.

Results: : 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.

Conclusions: : 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.

Keywords: imaging/image analysis: non-clinical • ganglion cells • synapse 
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