July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
OFF-layer Branches of ON Cone Bipolar Cells in Early Retinal Degeneration
Author Affiliations & Notes
  • Jessica Garcia
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Rebecca L Pfeiffer
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Crystal Sigulinsky
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • James Anderson
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Daniel Emrich
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Jeebika Dahal
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Hope Morrison
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Kevin Rapp
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Jia-Hui Yang
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Carl Watt
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Mineo Kondo
    Mie University, Graduate School of Med, Tsu, Japan
  • Hiroko Terasaki
    Nagoya University, School of Medicine, Nagoya-shi, Japan
  • Robert Marc
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Bryan W Jones
    Ophthamology, Moran Eye Center University of Utah, Salt Lake City, Utah, United States
  • Footnotes
    Commercial Relationships   Jessica Garcia, None; Rebecca Pfeiffer, None; Crystal Sigulinsky, None; James Anderson, None; Daniel Emrich, None; Jeebika Dahal, None; Hope Morrison, None; Kevin Rapp, None; Jia-Hui Yang, None; Carl Watt, None; Mineo Kondo, None; Hiroko Terasaki, None; Robert Marc, Signature Immunologics Inc. (I); Bryan Jones, None
  • Footnotes
    Support  R01 Grant EY015128, R01 Grant EY02576, T32 Grant EY024234, P30 Grant EY014800, Undergraduate research awards from the Undergraduate Research Opportunities Program at the University of Utah, Unrestricted Grant from Research to Prevent Blindness to the Department of Ophthalmology & Visual Sciences
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 555. doi:https://doi.org/
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      Jessica Garcia, Rebecca L Pfeiffer, Crystal Sigulinsky, James Anderson, Daniel Emrich, Jeebika Dahal, Hope Morrison, Kevin Rapp, Jia-Hui Yang, Carl Watt, Mineo Kondo, Hiroko Terasaki, Robert Marc, Bryan W Jones; OFF-layer Branches of ON Cone Bipolar Cells in Early Retinal Degeneration. Invest. Ophthalmol. Vis. Sci. 2019;60(9):555. doi: https://doi.org/.

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

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Abstract

Purpose : Cone bipolar cells are customarily classified into superclasses of ON (ON-BCs) and OFF (OFF-BCs). ON or OFF specialization is further segregated by stratification within the inner plexiform layer. Retinal degeneration induces negative plasticity termed remodeling, that includes aberrant neurite extension from multiple cell types (rewiring) and ON-BCs switching their glutamate receptors to match that of OFF-BCs (reprogramming). Previous analysis in healthy retina shows that ON-BCs can make small ribbon synapses in the descending axon or, less frequently, simple single branch projections in the OFF layer. What impact remodeling has on these OFF-layer branches is unknown. In this study, we compare OFF branches from ON-BCs in a connectome of early retinal degeneration (RPC1) to our healthy retinal connectome (RC1).

Methods : Retinal tissues selected for RC1 and RPC1 were collected post-mortem from a 13 month old Dutch-belted healthy female rabbit and a male 10 month old transgenic P347L rabbit model of autosomal dominant retinitis pigmentosa, respectively. RPC1 shows signs of remodeling including rod outer-segment degeneration and aberrant neurite extension. Tissues were fixed in mixed aldehydes and subsequently osmicated, dehydrated, resin embedded, and sectioned at 90 nm (RC1) or 70 nm (RPC1). Sections were placed on formvar grids, stained, and imaged at 2nm/px on a JEOL JEM-1400 TEM using SerialEM software. 1 section was reserved from every 30 for Computational Molecular Phenotyping, and probed for small molecules. Both volumes were evaluated using the Viking software suite.

Results : Ribbons of ON-BCs formed in the OFF layer branches have been previously described to contact glycinergic amacrine cells (ACs), GABAergic ACs, ON ganglion cells, and intrinsically photosensitive ganglion cells. Initial analysis of OFF branches of ON-BCs in RPC1 demonstrate more complex branching than in RC1 and increased number of synapses on these branches. In contrast to the inconsistent OFF layer branch stratification observed in RC1, the OFF branches in RPC1 appear to stratify at a similar level. Evaluation of synaptic partners is ongoing.

Conclusions : Increased complexity and number of synapses found in the OFF branches of some ON-BCs ultimately may represent ON network corruption. Exploring synaptic partners will reveal potential network alterations in retinal degenerative disease.

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

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