Experimental setup compatible with anesthetized/awake rats and data acquisition settings

We established experimental setups that accommodated both anesthetized and awake rats. The schematics of the implemented wearable tFUS headgear, with the transducer actuation systems, are shown in Fig. 1d. A swivel connector (slip ring with flange-736; Adafruit, New York, NY) was located above the middle of the cage/platform, granting unrestricted motion and access to a power source for actuating the transducer during the awake tFUS sessions. A data acquisition system (PowerLab 8/30 and LabChart 7; ADInstruments, Colorado Springs, CO) was used to acquire time-series data of sonication events (onset timing and duration), being synchronized with a video recording (29.97 frames per second; FPS, by QTH44; Q-See; Anaheim, CA) to analyze the location and onset timing of the movement elicited by the sonication. Additionally, a light-emitting diode (LED), turned on in-sync with each sonication event, was placed within the field-of-view of the video recording as a visual indicator of the sonication timing (shown in Fig. 2a–c, upper panels).

The experimental sessions (upper panels) and the merged images before/after tail movement (lower panels). a Freely-moving awake rats, as well as under light anesthesia of b ketamine/xylazine, or c isoflurane. The location of LED that shows the timing and duration of sonication is shown in dotted red circles. The movement onset (‘Mov onset’) latencies with respect to the FUS onset are also shown in the lower panels. The arrows indicate the elicited movement (see Additional files 1, 2, 3)