Two-photon optogenetics-based assessment of neuronal connectivity in healthy and chronic hypoperfusion mice

Masaki Yoshioka; Manami Takahashi; Jeff Kershaw; Mariko Handa; Ayaka Takada; Hiroyuki Takuwa

doi:10.1117/1.NPh.11.3.035009

28 September 2024 Two-photon optogenetics-based assessment of neuronal connectivity in healthy and chronic hypoperfusion mice

Masaki Yoshioka, Manami Takahashi, Jeff Kershaw, Mariko Handa, Ayaka Takada, Hiroyuki Takuwa

Author Affiliations +

Neurophotonics, Vol. 11, Issue 3, 035009 (September 2024). https://doi.org/10.1117/1.NPh.11.3.035009

Abstract

Significance

Two-photon optogenetics and simultaneous calcium imaging can be used to visualize the response of surrounding neurons with respect to the activity of an optically stimulated target neuron, providing a direct method to assess neuronal connectivity.

Aim

We aim to develop a two-photon optogenetics-based method for evaluating neuronal connectivity, compare it to the existing indirect resting-state synchrony method, and investigate the application of the method to brain pathophysiology.

Approach

C1V1-mScarlet was introduced into GCaMP6s-expressing transgenic mice with an adeno-associated virus. Optical stimulation of a single target neuron and simultaneous calcium imaging of the target and surrounding cells were performed. Neuronal connectivity was evaluated from the correlation between the fluorescence intensity of the target and surrounding cells.

Results

The neuronal connectivity in the living brain was evaluated using two-photon optogenetics. However, resting-state synchrony was not always consistent with two-photon optogenetics-based connectivity. Comparison with neuronal synchrony measured during sensory stimulation suggested that the disagreement was due to external sensory input. Two-photon optogenetics-based connectivity significantly decreased in the common carotid artery occlusion model, whereas there was no significant change in the control group.

Conclusions

We successfully developed a direct method to evaluate neuronal connectivity in the living brain using two-photon optogenetics. The technique was successful in detecting connectivity impairment in hypoperfusion model mice.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 International License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

Citation Download Citation

Masaki Yoshioka, Manami Takahashi, Jeff Kershaw, Mariko Handa, Ayaka Takada, and Hiroyuki Takuwa "Two-photon optogenetics-based assessment of neuronal connectivity in healthy and chronic hypoperfusion mice," Neurophotonics 11(3), 035009 (28 September 2024). https://doi.org/10.1117/1.NPh.11.3.035009

Received: 6 March 2024; Accepted: 22 August 2024; Published: 28 September 2024

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