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Published December 15, 2023 | v1
Journal Article Open

Effects of focused ultrasound in a "clean" mouse model of ultrasonic neuromodulation


Recent studies on ultrasonic neuromodulation (UNM) in rodents have shown that focused ultrasound (FUS) can activate peripheral auditory pathways, leading to off-target and brain-wide excitation, which obscures the direct activation of the target area by FUS. To address this issue, we developed a new mouse model, the double transgenic Pou4f3^(+/DTR) × Thy1-GCaMP6s, which allows for inducible deafening using diphtheria toxin and minimizes off-target effects of UNM while allowing effects on neural activity to be visualized with fluorescent calcium imaging. Using this model, we found that the auditory confounds caused by FUS can be significantly reduced or eliminated within a certain pressure range. At higher pressures, FUS can result in focal fluorescence dips at the target, elicit non-auditory sensory confounds, and damage tissue, leading to spreading depolarization. Under the acoustic conditions we tested, we did not observe direct calcium responses in the mouse cortex. Our findings provide a cleaner animal model for UNM and sonogenetics research, establish a parameter range within which off-target effects are confidently avoided, and reveal the non-auditory side effects of higher-pressure stimulation.

Copyright and License

© 2023 The Author(s). This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).


The authors thank Joseph Wekselblatt, David Mittelstein, and Pierina Barturen-Larrea for helpful discussion, and members of the Shapiro and Tsao labs for assistance with experiments. This research was supported by NIH BRAIN Initiative grant R24MH106107 (Co-PIs D.Y.T. and M.G.S.), RF1MH117080 (to M.G.S.), and the Howard Hughes Medical Institute (D.Y.T.). Related research in the Shapiro Laboratory is also supported by the Howard Hughes Medical Institute and the Packard Fellowship in Science and Engineering.


H.G., D.Y.T., and M.G.S. conceived the study. H.G. and M.G.S. designed all experiments, and H.G. performed all the in vivo experiments and analyzed the data. D.W. and S.J.Y. performed the in vitro experiments in cell cultures and analyzed the data. H.S. performed the modeling. T.S. and M.G.S. initialized the idea of using the double transgenic mouse model. H.G. and M.G.S. wrote the manuscript with input from all authors.

Conflict of Interest

The authors declare no competing interests.

Additional Information

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December 4, 2023
December 13, 2023