Circuit mechanisms underlying embryonic retinal waves

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Abstract

Spontaneous activity is a hallmark of developing neural systems. In the retina, spontaneous activity comes in the form of retinal waves, comprised of three stages persisting from embryonic day 16 (E16) to eye opening at postnatal day 14 (P14). Though postnatal retinal waves have been well characterized, little is known about the spatiotemporal properties or the mechanisms mediating embryonic retinal waves, designated Stage 1 waves. Using a custom-built macroscope to record spontaneous calcium transients from whole embryonic retinas, we show that Stage 1 waves are initiated at several locations across the retina and propagate across a broad range of areas. Blocking gap junctions reduced the frequency and size of Stage 1 waves, nearly abolishing them. Global blockade of nAChRs similarly nearly abolished Stage 1 waves. Thus, Stage 1 waves are mediated by a complex circuitry involving subtypes of nAChRs and gap junctions. Stage 1 waves in mice lacking the β2 subunit of the nAChRs (β2-nAChR-KO) persisted with altered propagation properties and were abolished by a gap junction blocker. To assay the impact of Stage 1 waves on retinal development, we compared the spatial distribution of a subtype of retinal ganglion cells, intrinsically photosensitive retinal ganglion cells (ipRGCs), which undergo a significant amount of cell death, in WT and β2-nAChR-KO mice. We found that the developmental decrease of ipRGC density is preserved between WT and β2-nAChR-KO mice, indicating that processes regulating ipRGC distribution are not influenced by spontaneous activity.

Data availability

We have uploaded the raw data for Figure 5 on Dryad.All other raw imaging data and images are available upon request as they are too large to update to on Dryad. They are residing on our lab server and can be transferred via ftp.Figures 1-4. These data are based on movies acquired from live imaging of activity using a macroscope or a 2-photon scanning microscope.Figure 1: 50 gigabytesFigures 2/4: 180 gigabytesFigure 3: 233 gigabytesFigure 5: These are high resolution fluorescence images acquired from microscope at various z-plane focus planes. 7 gigabytes total. (on Dryad)All code and software are available in this gitHub location: https://github.com/FellerLabCodeShare/Embryonic-Retinal-WavesInstructions on how to build a macroscope available at this gitHub location: https://github.com/Llamero/DIY_Epifluorescence_Macroscope

The following data sets were generated

1. Voufo C
2. Chen AQ
3. Smith B
4. Yan R
5. Feller MB
6. Tiriac A
(2023) Circuit mechanisms underlying embryonic retinal waves
Dryad Digital Repository, doi:10.5061/dryad.h18931zr2.

https://dx.doi.org/10.5061/dryad.h18931zr2

Article and author information

Author details

Christiane Voufo

Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, United States

Competing interests
No competing interests declared.
Andy Quaen Chen

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0002-4970-045X
Benjamin E Smith

School of Optometry, University of California, Berkeley, Berkeley, United States

Competing interests
No competing interests declared.
Rongshan Yan

Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, United States

Competing interests
No competing interests declared.
Marla B Feller

Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

For correspondence
mfeller@berkeley.edu

Competing interests
Marla B Feller, Reviewing editor, eLife.

"This ORCID iD identifies the author of this article:" 0000-0002-9137-5849
Alexandre Tiriac

Biological Sciences, Vanderbilt University, Nashville, United States

For correspondence
alexandre.tiriac@vanderbilt.edu

Competing interests
No competing interests declared.

Funding

National Eye Institute (RO1EY013528,RO1EY019498,P30EY003176)

Christiane Voufo

National Eye Institute (K99EY030909)

Alexandre Tiriac

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication

Ethics

Animal experimentation: All animal procedures were approved by the UC Berkeley Institutional Animal Care and Use Committee and conformed to the NIH Guide for the Care and Use of Laboratory Animals, the Public Health Service Policy, and the SFN Policy on the Use of Animals in Neuroscience Research.

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.