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Optimized ultrasound neurostimulation of ventromedial prefrontal cortex reverses depression behaviours and normalizes brain metabolism

Marc Legrand, Laurent Galineau, Anthony Novell, Barbara Planchez, Bruno Brizard, Samuel Leman, Clovis Tauber, Jean-Michel Escoffre, Sophie Serriere, Tarik Iazourene, Wissam El-Hage, Patrick Emond, Catherine Belzung, Ayache Bouakaz

npj Acoustics 2026, 2 · 10.1038/s44384-026-00050-z

rodentdepressionanxietybehaviourpethistology molecularemg mepother
Speciesmouse (BALB/cByJRj)
Subjects30, 15swept animals
Sessions per subject1, 5swept
Randomisedyes
Blindingsingle
Sham / controlundescribed
Auditory controlnot reported
Readout timingboth
Anaesthesiaanaesthetised
Readoutsbehaviour, pet, histology molecular, emg mep, other18F-FDG microPET; c-Fos immunohistochemistry; targeted LC-HRMS metabolomics; electromyographic motor-evoked potentials used for pressure-response optimisation
Direction of effectexcitatoryRepeated IL-USNS increased local c-Fos expression in the infralimbic cortex (target engagement) and reversed UCMS-induced deficits in nest-building and anxiety-like open-field exploration, with sustained hypermetabolism (FDG-PET) in vmPFC/IL and connected limbic regions.
Adverse eventsnone observedFurthermore, metabolomic analysis revealed no increase in inflammatory or excitotoxic markers typically associated with tissue damage<sup>54,55</sup>, supporting an excellent safety profile.

Exposures

Exposure 1: Single-pulse pressure-response mapping at primary motor cortex (M1)

Target: primary motor cortex — “M1 (primary motor cortex)
Device: IGT / Imasonic · Imasonic

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)160✓✓
Pulse repetition frequency (Hz)0.1✓✓
Duty cycle (%)1.6pulse duration × PRF gives 1.6%✓✓
Sonication duration (s)0.16✓✓
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 estimatederatingmean or range across subjects
In-situ pressure (kPa)50, 500swept✓✓
In-situ Isppa (W/cm²)5.33✓✓
In-situ Ispta (W/cm²)0.085✓✓
Protocol, in the paper’s words

Single continuous 160 ms pulses delivered to left M1 at pressures stepped from 50 to 500 kPa in 50 kPa increments, ten pulses per step, repeated every 10 s (0.1 Hz); this inter-pulse repetition corresponds to a stated 1.6% duty cycle across the pulse train (distinct from the internal waveform, which is a single continuous burst per pulse). Motor success (visible forepaw twitch, n=20) and motor-evoked potential amplitude (EMG, n=10) were used to derive a motor threshold of 250 kPa and an optimal plateau at 400 kPa.

Exposure 2: Repeated-pulse USNS of the infralimbic cortex (IL)

Target: ventromedial prefrontal cortex — “infralimbic cortex (IL), rodent homologue of vmPFC
Device: IGT / Imasonic · Imasonic

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)160✓✓
Pulse repetition frequency (Hz)0.1✓✓
Duty cycle (%)1.6pulse duration × PRF gives 1.6%✓✓
Sonication duration (s)600✓✓
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 estimatederatingsingle value
In-situ pressure (kPa)400✓✓
In-situ Isppa (W/cm²)5.33✓✓
In-situ Ispta (W/cm²)0.085✓✓
Protocol, in the paper’s words

60 single 160 ms pulses at 400 kPa delivered at 0.1 Hz (every 10 s) over a 10 min session. For focality confirmation (c-Fos), one such session was given to 4 active mice (vs 4 sham-anaesthesia mice); for the chronic UCMS experiment, one session per day was repeated for 5 consecutive days (days 29-33), targeted stereotaxically at bregma +2.0 mm, midline.

Flags from extraction

  • n_subjectsGiven as group-size list per instructions rather than summed: 30 mice for parameter-setting (M1 motor screening n=20 plus EMG n=10, a subset of which were later reused for the 8-mouse IL c-Fos focality experiment), and 15 mice in the chronic IL-USNS treatment arm of the UCMS cohort; other UCMS arms (sham-USNS, vehicle, fluoxetine, non-stressed) did not receive ultrasound.
  • sham_type'Sham-USNS'/'sham controls' are described only as anaesthesia without ultrasound; the paper does not state whether a transducer was placed but powered off or simply absent, so the sham mechanism is not fully described.
  • exposures[0].timing.duty_cycle_pctText states 'duty cycle 1.6%' for the repeated 160 ms pulses at 0.1 Hz, but this describes the inter-pulse repetition duty cycle across a pulse train, not the internal pulse waveform (which is a single continuous burst); left null per the continuous-wave rule and described in protocol_description instead.
  • n_sessions_per_subjectVaries by experiment: single session for M1 optimisation and the 8-mouse c-Fos focality test, versus 5 consecutive daily sessions for the chronic UCMS IL-USNS group.
  • exposures[0].timing.waveformset to pulsed by the tie-break: the ruled timing has a pulse repetition or duty cycle
  • exposures[1].timing.waveformset to pulsed by the tie-break: the ruled timing has a pulse repetition or duty cycle