← Explore

Manipulation of Subcortical and Deep Cortical Activity in the Primate Brain Using Transcranial Focused Ultrasound Stimulation

Davide Folloni, Lennart Verhagen, Rogier B. Mars, Elsa Fouragnan, Charlotte Constans, Jean-François Aubry, Matthew F.S. Rushworth, Jérôme Sallet

Neuron 2019, 101, 1109-1116.e5 · 10.1016/j.neuron.2019.01.019

nonhuman primatehealthyfmri

Abstract

The causal role of an area within a neural network can be determined by interfering with its activity and measuring the impact. Many current reversible manipulation techniques have limitations preventing their application, particularly in deep areas of the primate brain. Here, we demonstrate that a focused transcranial ultrasound stimulation (TUS) protocol impacts activity even in deep brain areas: a subcortical brain structure, the amygdala (experiment 1), and a deep cortical region, the anterior cingulate cortex (ACC, experiment 2), in macaques. TUS neuromodulatory effects were measured by examining relationships between activity in each area and the rest of the brain using functional magnetic resonance imaging (fMRI). In control conditions without sonication, activity in a given area is related to activity in interconnected regions, but such relationships are reduced after sonication, specifically for the targeted areas. Dissociable and focal effects on neural activity could not be explained by auditory confounds.

Abstract via europepmc.

Speciesrhesus macaque (Macaca mulatta)
Subjects4, 3swept animals
Sessions per subjectnot reported
Randomisedno
Blindingnone
Sham / controlnone
Auditory controlother
Readout timingoffline
Anaesthesiaanaesthetised
ReadoutsfmriResting-state fMRI (BOLD) functional connectivity / 'connectivity fingerprint' analysis between stimulated region and rest of brain
Direction of effectnot assessedTUS reduced resting-state functional coupling (BOLD connectivity) between the stimulated area (amygdala or ACC) and its normally interconnected network, specifically for the stimulated site; this is a change in functional connectivity, not a direct measure of increased/decreased neural activity, so direction of the underlying excitatory/inhibitory effect was not assessed.
Adverse eventsnone observedStructural MRI scans collected shortly after TUS in the present study showed no evidence of transient edema (Figure S4).

Exposures

Exposure 1: Experiment 1: amygdala TUS

Target: amygdala — “amygdala
Device: Sonic Concepts · Sonic Concepts, Bothell, WA, USA · H115-MR (64 mm diameter, 51.74 mm focal depth, 250 kHz)

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)250✓✓
Pulse duration (ms)30✓✓
Pulse repetition frequency (Hz)10✓✓
Duty cycle (%)30pulse duration × PRF gives 30%✓✓
Sonication duration (s)40✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)1,150, 1,270swept✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatesimulationsingle value
In-situ pressure (kPa)1,440✓✓
In-situ Isppa (W/cm²)64.9✓✓
In-situ Ispta (W/cm²)19.5✓✓
Protocol, in the paper’s words

A single 40-s train of pulsed ultrasound (250 kHz) comprising 30-ms bursts every 100 ms (duty cycle 30%) was directed to the target region using a single-element transducer with a region-specific water-filled coupling cone; recordings of neural (fMRI) activity began ~30 min after the end of the 40-s TUS train ('offline' protocol).

Exposure 2: Experiment 2: ACC TUS

Target: anterior cingulate cortex — “anterior cingulate cortex (ACC)
Device: Sonic Concepts · Sonic Concepts, Bothell, WA, USA · H115-MR (64 mm diameter, 51.74 mm focal depth, 250 kHz)

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)250✓✓
Pulse duration (ms)30✓✓
Pulse repetition frequency (Hz)10✓✓
Duty cycle (%)30pulse duration × PRF gives 30%✓✓
Sonication duration (s)40✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)1,150, 1,270swept✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatesimulationsingle value
In-situ pressure (kPa)780✓✓
In-situ Isppa (W/cm²)18.8✓✓
In-situ Ispta (W/cm²)5.63✓✓
Protocol, in the paper’s words

A single 40-s train of pulsed ultrasound (250 kHz) comprising 30-ms bursts every 100 ms (duty cycle 30%) was directed to the target region using a single-element transducer with a region-specific water-filled coupling cone; recordings of neural (fMRI) activity began ~30 min after the end of the 40-s TUS train ('offline' protocol).

Flags from extraction

  • exposures[0].free_field.pressure_kpaFree-field pressure (1.15-1.27 MPa measured in water) is reported as a general calibration range across the study, not broken out separately per target; midpoint (1.21 MPa) recorded per exposure and applied identically to both experiments.
  • auditory_controlNo masking/deafening procedure was used; instead the carrier (250 kHz) and pulse-envelope/PRF (10 Hz) were deliberately chosen to fall outside the macaque audible range, and neural recording began ~30 min after TUS ended. Classified as 'other' since no listed category matches this design-based control.
  • randomisedPaper states all available animals took part with no pre-selection or restriction on group allocation, implying non-random, opportunistic assignment; not explicitly labelled 'randomised' or 'non-randomised' by the authors.

Notes: Two exposures correspond to Experiment 1 (amygdala, n=4) and Experiment 2 (ACC, n=3); an additional n=9 animals served as no-stimulation controls (some overlap with the 11 total animals). In-situ (simulated, finite-element head model) pressure/intensity values are target-specific and taken directly from the text; free-field (water-measured) pressure is a general per-study calibration range.