July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Axon-bearing amacrine cells labelled in transgenic GlyT2 mice.
Author Affiliations & Notes
  • Ben Sivyer
    Ophthalmology, Casey Eye Institute, Oregon Health & Sciences Univ, Portland, Oregon, United States
  • Marc Andrew Meadows
    Vollum Institute, Oregon Health & Science University, Portland, Oregon, United States
  • Patrick C Kerstein
    Vollum Institute, Oregon Health & Science University, Portland, Oregon, United States
  • Kevin M Wright
    Vollum Institute, Oregon Health & Science University, Portland, Oregon, United States
    Ophthalmology, Casey Eye Institute, Oregon Health & Sciences Univ, Portland, Oregon, United States
  • Henrique Von Gersdorff
    Vollum Institute, Oregon Health & Science University, Portland, Oregon, United States
    Ophthalmology, Casey Eye Institute, Oregon Health & Sciences Univ, Portland, Oregon, United States
  • Footnotes
    Commercial Relationships   Ben Sivyer, None; Marc Meadows, None; Patrick Kerstein, None; Kevin Wright, None; Henrique Von Gersdorff, None
  • Footnotes
    Support  Bright Focus Foundation, Unrestricted grant from Research to Prevent Blindness.
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 542. doi:
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    • Get Citation

      Ben Sivyer, Marc Andrew Meadows, Patrick C Kerstein, Kevin M Wright, Henrique Von Gersdorff; Axon-bearing amacrine cells labelled in transgenic GlyT2 mice.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):542.

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

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Abstract

Purpose : Approximately half of the total population of retinal amacrine cells are glycinergic yet we have little knowledge of the functional roles of most glycinergic ACs in visual processing. To address this we crossed GlyT2-cre mouse (GlyT2; SL6A5) with Ai9 floxed tdTomato reporter line (Jax007909).

Methods : We made isolated retinal whole-mount preparations from Glyt2-cre x Ai9 mice and GlyT2 EGFP mice. TdTomato and EGFP positive neurons in the retinal ganglion cell layer were target for electrophysiological recordings or Neurobiotin electroporation. Post-fixed retinas were imaged with confocal microscopy to determine neuronal morphology and immunohistochemical co-localization with antibodies for RPBMS, GlyT1/2 and nNOS.

Results : GlyT2-cre; tdTomato mice have labeled neurons in both the inner nuclear layer (INL; 2894 to 4615 cells/mm) and retinal ganglion cell layer (GCL; 290 to 419 cells/mm2 n =4 ), where 4.2 ± 2.6% are labelled with an antibody to RPBMS indicating a subpopulation of these cells are retinal ganglion cells. A further subpopulation of GlyT2 amacrine cells in the RGC layer (3.9 ± 0.8%) were immunopositive for nNOS but none were immunopositive for Gly-T1/2. 79.1 ± 7.7% of INL tdTomato positive cells were immunopositive for GlyT1/2 indicating this transgenic line might also label GABAergic amacrine cells. Physiological recordings were predominantly from spiking amacrine cells (n = 8 from 4 mice). Imaging after Neurobiotin electroporation confirmed these were wide-field amacrine cells with asymmetrical dendritic arbors approximately 100 um in diameter and wide-ranging axonal arbors projecting millimeters across the retina. The dendritic arbors of tdTomato positive WF ACs were interspersed with large (4.6 ± 1.2 µm2) basket-like varicosities and dendritic spines (2.59 ± 1.13 µm spine length). The most commonly encountered RGC layer amacrine cell type from Glyt2-cre;tdTomato mice depolarize in response to light flash stimuli and fired a sustained burst of action potentials.

Conclusions : Our results suggest GlyT2-cre transgenic mice label a mixed population of glycinergic and GABAergic neurons but are useful for selectively targeting axon-bearing amacrine cells in the GCL of the mouse retina.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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