June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Intersectional strategies for targeting amacrine and ganglion cell types in the mouse retina
Author Affiliations & Notes
  • Yongling Zhu
    Northwestern University, Chicago, Illinois, United States
  • Jian Xu
    Northwestern University, Chicago, Illinois, United States
  • Hongkui Zeng
    Allen Institute for Brain Science, Seattle, Washington, United States
  • Steven H DeVries
    Northwestern University, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Yongling Zhu, None; Jian Xu, None; Hongkui Zeng, None; Steven DeVries, None
  • Footnotes
    Support  NIH Grant EY018204, Research to Prevent Blindness
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 239. doi:
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      Yongling Zhu, Jian Xu, Hongkui Zeng, Steven H DeVries; Intersectional strategies for targeting amacrine and ganglion cell types in the mouse retina. Invest. Ophthalmol. Vis. Sci. 2017;58(8):239.

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

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Abstract

Purpose : Systematic access to amacrine and ganglion cell types will facilitate the study of the organization and function of inner retinal circuits. Current genetic approaches exploit the molecular diversity of neurons by driving Cre expression with specific gene promoters. However, Cre expression is rarely confined to a single cell type. Strategies for restricting transgene expression to a single neuron type are essential for obtaining unambiguous information about processing. We employ an intersectional strategy based on combined response in a single cell type to expression driven by two distinct gene promoters.

Methods : We created various intersections among two binary systems (Cre/loxP and Flp/FRT) in which Cre and Flp expression are controlled by different promoters, coupled with reporter genes that can be activated only in the cells that contain both Cre and Flp. Specifically, we characterized six newly developed Flp driver lines, followed by crossing them with Cre driver lines that are driven by different promoters, but which show overlap with Flp lines in expression patterns. Once the Flp driver line and Cre driver line were crossed, Cre and Flp double–dependent reporter mouse lines were introduced, allowing specific labelling of the cells within the Cre and Flp intersection. The morphological characteristics of the labeled cells, including soma location, dendritic tree size, and the stratification level were analyzed using standard confocal imaging and landmarks in ChAT labeled tissue. Cell types were identified based on published work.

Results : We have created more than 20 combinational mouse lines through selective cross breeding of ~20 Cre driver lines, 6 Flp driver lines and corresponding reporter lines. We demonstrate that, with appropriate combinations between Cre and Flp drivers, the specificity of cell type targeting is greatly enhanced.

Conclusions : Intersectional strategies, by providing a genetic solution to restrict transgene expression to specific retinal cell types, can accelerate progress in exploring the functional operation of inner retinal circuits.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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