Prior to the behavior test, mice were adapted to the behavior recording environment by placing the mouse in the behavior testing arena with the power amplifier turned on. During the behavior recording, mice were lightly anesthetized with isoflurane (1% induction and maintenance). The base plate on the mouse and the wearable ultrasound transducer were both sufficiently filled with degassed ultrasound gel (Aquasonics). The wearable transducer was then securely plugged into the base plate of the mouse, and the mouse was then placed in a circular arena on a heating pad for 30 min to allow the mouse body temperature to recover from any possible anesthesia effects. The heating pad was then removed, and the mouse was allowed to habituate for 15 min in the actual behavior test arena.
During the recording period, FUS was applied at a frequency of 1.5 MHz, duty cycle (DC) of 40%, pulse repetition frequency (PRF) of 10 Hz, and 15 s total sonication duration with 185 s inter-stimulation interval (ISI) for a total of five stimulations. The onset and offset of the ultrasound pulse was smoothed to avoid possible auditory effects [29 (link)]. The acoustic pressures used in the study were 0, 0.7, and 1.1 MPa (measured in a water tank) to investigate the effect of pressure on locomotor behavior outcomes. These parameters corresponded to mechanical indices of 0, 0.57, and 0.90 (in water), respectively. Custom MATLAB software was used to control when ultrasound was applied via an Arduino Uno. A red LED attached to the Arduino Uno would turn on when ultrasound was applied to precisely synchronize mouse behavior to each FUS stimulation. In each group, mice were given five consecutive FUS stimulations at one pressure.
During the recording period, FUS was applied at a frequency of 1.5 MHz, duty cycle (DC) of 40%, pulse repetition frequency (PRF) of 10 Hz, and 15 s total sonication duration with 185 s inter-stimulation interval (ISI) for a total of five stimulations. The onset and offset of the ultrasound pulse was smoothed to avoid possible auditory effects [29 (link)]. The acoustic pressures used in the study were 0, 0.7, and 1.1 MPa (measured in a water tank) to investigate the effect of pressure on locomotor behavior outcomes. These parameters corresponded to mechanical indices of 0, 0.57, and 0.90 (in water), respectively. Custom MATLAB software was used to control when ultrasound was applied via an Arduino Uno. A red LED attached to the Arduino Uno would turn on when ultrasound was applied to precisely synchronize mouse behavior to each FUS stimulation. In each group, mice were given five consecutive FUS stimulations at one pressure.
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