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

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posted on 2023-06-09, 06:58 authored by Manoj Kulkarni, Timm Schubert, Thomas BadenThomas Baden, Bernd Wissinger, Thomas Euler, Francois Paquet-Durand
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.

History

Publication status

  • Published

File Version

  • Published version

Journal

Journal of Visualized Experiments

ISSN

1940-087X

Publisher

JoVE

Issue

99

Article number

e52588

Department affiliated with

  • Neuroscience Publications

Research groups affiliated with

  • Sussex Neuroscience Publications

Full text available

  • Yes

Peer reviewed?

  • Yes

Legacy Posted Date

2017-07-03

First Open Access (FOA) Date

2017-07-03

First Compliant Deposit (FCD) Date

2017-07-03

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