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Ultrasonic Neuromodulation Causes Widespread Cortical Activation via an Indirect Auditory Mechanism

Tomokazu Sato, Mikhail G. Shapiro, Doris Y. Tsao

Neuron 2018, 98, 1031-1041.e5 · 10.1016/j.neuron.2018.05.009

rodenthealthycellular imagingemg mep

Abstract

Ultrasound has received widespread attention as an emerging technology for targeted, non-invasive neuromodulation based on its ability to evoke electrophysiological and motor responses in animals. However, little is known about the spatiotemporal pattern of ultrasound-induced brain activity that could drive these responses. Here, we address this question by combining focused ultrasound with wide-field optical imaging of calcium signals in transgenic mice. Surprisingly, we find cortical activity patterns consistent with indirect activation of auditory pathways rather than direct neuromodulation at the ultrasound focus. Ultrasound-induced activity is similar to that evoked by audible sound. Furthermore, both ultrasound and audible sound elicit motor responses consistent with a startle reflex, with both responses reduced by chemical deafening. These findings reveal an indirect auditory mechanism for ultrasound-induced cortical activity and movement requiring careful consideration in future development of ultrasonic neuromodulation as a tool in neuroscience research.

Abstract via europepmc.

Speciesmouse (Thy1-GCaMP6s transgenic; C57BL/6J)
Subjects10, 8, 7, 5swept animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlnone
Auditory controldeafened subjects, sound only sham
Readout timingboth
Anaesthesiaanaesthetised
Readoutscellular imaging, emg mepWide-field cortical calcium imaging (GCaMP6s); hindlimb EMG
Direction of effectmixed or unclearUltrasound produced strong, early excitatory activation of the contralateral auditory cortex closely resembling responses to audible sound, whereas the directly targeted visual cortex showed only mixed weak inhibitory/delayed excitatory signals with no clear evidence of direct neuromodulation. Motor (EMG) responses to ultrasound resembled a startle reflex and were greatly reduced by chemical deafening, implicating an indirect auditory pathway rather than direct cortical neuromodulation.
Adverse eventsnot reported

Exposures

Exposure 1: Focused ultrasound to posterior visual cortex, 5 intensities

Target: visual cortex — “posterior portion of the visual cortex
Device: Blatek · Blatek · AT24020

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)0.2✓✓
Pulse repetition frequency (Hz)1,500✓✓
Duty cycle (%)30pulse duration × PRF gives 30%✓✓
Sonication duration (s)0.08✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatenot reported
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Ispta, domain unspecified (W/cm²)4.2, 1.3, 0.38, 0.11, 0.034swept✓✓
Protocol, in the paper’s words

All ultrasound stimuli were of 500 kHz acoustic frequency, 100 cycles/pulse, 1.5 kHz pulse repetition frequency, and 120 pulses, yielding a duty cycle of 30% and a stimulus duration of approximately 80 ms. Each imaging block presented one trial each of 5 ultrasound intensities, 4 sound intensities, and a light flash in random order (200 blocks per imaging session); EMG sessions used 25 trials per stimulus type (sound, ultrasound, air puff, light) with a 10 s inter-trial interval, in two blocks 30 and 150 min after final injection.

Flags from extraction

  • exposures[0].timing.pulse_duration_msText states pulse duration of 200 ms, but 100 cycles/pulse at 500 kHz and a duty cycle of 30% at a PRF of 1500 Hz arithmetically imply a pulse duration of ~0.2 ms (200 microseconds); recorded as stated in the text.
  • exposures[0].unspecified_domain.ispta_w_cm2Methods list the ultrasound intensity series as 4.2, 1.3, 0.38, 0.11, 0.034 W/cm2, but the Figure 4C legend gives the second-highest value as 1.4 W/cm2; recorded the Methods values.
  • exposures[0].unspecified_domainPaper labels these values I_SPTA in a figure legend but does not state whether intensities used for stimulation were measured in free field (water tank) or at the target through the (thinned) skull; placed in unspecified_domain.
  • n_subjectsNo single total N is stated; recorded as the four distinct group sizes reported across the imaging cohort (n=10) and the three EMG deafening-experiment cohorts (deafened n=8, saline n=7, no-gel n=5).
  • sham_typeNo sham/inactive-ultrasound arm was used; the comparisons are across stimulus modalities (ultrasound, sound, light, air puff), so sham_type is recorded as none.