Imaging Ca²+ dynamics in cone photoreceptor axon terminals of the mouse retina

Kulkarni, Manoj, Schubert, Timm, Baden, Tom, Wissinger, Bernd, Euler, Thomas and Paquet-Durand, Francois (2015) Imaging Ca²+ dynamics in cone photoreceptor axon terminals of the mouse retina. Journal of Visualized Experiments (99). e52588. ISSN 1940-087X

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Abstract

Retinal cone photoreceptors (cones) serve daylight vision and are the basis of color discrimination. They are subject to degeneration, often leading to blindness in many retinal diseases. Calcium (Ca2+), a key second messenger in photoreceptor signaling and metabolism, has been proposed to be indirectly linked with photoreceptor degeneration in various animal models. Systematically studying these aspects of cone physiology and pathophysiology has been hampered by the difficulties of electrically recording from these small cells, in particular in the mouse where the retina is dominated by rod photoreceptors. To circumvent this issue, we established a two-photon Ca2+ imaging protocol using a transgenic mouse line that expresses the genetically encoded Ca²+ biosensor TN-XL exclusively in cones and can be crossbred with mouse models for photoreceptor degeneration. The protocol described here involves preparing vertical sections (“slices”) of retinas from mice and optical imaging of light stimulus-evoked changes in cone Ca²+ level. The protocol also allows “in-slice measurement” of absolute Ca²+ concentrations; as the recordings can be followed by calibration. This protocol enables studies into functional cone properties and is expected to contribute to the understanding of cone Ca²+ signaling as well as the potential involvement of Ca²+ in photoreceptor death and retinal degeneration.

Item Type: Article
Schools and Departments: School of Life Sciences > Neuroscience
Research Centres and Groups: Sussex Neuroscience
Depositing User: Thomas Baden
Date Deposited: 03 Jul 2017 09:29
Last Modified: 03 Jul 2017 09:34
URI: http://sro.sussex.ac.uk/id/eprint/68926

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