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Efficacy of chronic ultrasound neurostimulation on behaviors and distributed brain metabolism in depressive-like mice

Marc Legrand, Laurent Galineau, Anthony Novell, Barbara Planchez, Bruno Brizard, Samuel Leman, Clovis Tauber, Jean-Michel Escoffre, Antoine Lefèvre, Philippe Gosset, Wissam El-Hage, Patrick Emond, Catherine Belzung, Ayache Bouakaz

2019 · 10.1101/813006

rodentdepressionanxietyemg mepbehaviourpethistology molecular

Abstract

Major depression is one of the main factors contributing to the Global Burden of Disease. Current treatment strategies ( e.g., antidepressants and neurostimulation techniques) of major depression show some limitations including inaccuracy and invasiveness. Ultrasound neurostimulation (USNS) has been recently introduced as a physical non-invasive method for brain tissue stimulation and has gained increasing interest. In this study, we sought to evaluate the efficacy of transcranial USNS in an unpredictable chronic mild stress (UCMS) mouse model. The results show that transcranial USNS of the infralimbic cortex reduced anxiety-related behaviors as well as some, but not all, depression-related parameters. [ 18 F]-FDG microPET imaging and brain metabolomic analyses showed that USNS triggered the activation of targeted brain region in addition to brain areas at a distance from the targeted zone, alleviating anxiety and depression-related behaviors induced by the UCMS regimen. Transcranial ultrasound neurostimulation show therapeutic potential in some aspects of major depression.

Abstract via europepmc.

Speciesmouse
Subjects20, 10, 4, 15swept animals
Sessions per subject1, 5swept
Randomisedyes
Blindingsingle
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingboth
Anaesthesiaboth
Readoutsemg mep, behaviour, pet, histology molecularElectrode-free video and EMG motor-evoked potentials (M1 parameter finding); c-Fos immunolabelling; [18F]-FDG microPET imaging; LC-HRMS brain metabolomics; nest-building, reward-maze and open-field behavioural tests
Direction of effectexcitatoryUSNS of M1 produced dose-dependent motor-evoked responses; acute/chronic USNS of the infralimbic cortex increased local and distant (striatum, thalamus, hippocampus, PAG, raphe) c-Fos expression and FDG metabolism, reduced anxiety-related behaviours (open-field mobility/center-crossings, nest-building) but did not improve anhedonia (reward-maze) relative to fluoxetine.
Adverse eventsnot reported

Exposures

Exposure 1: single-pulse USNS of primary motor cortex M1 for parameter optimization

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

Pulse timing
Waveformcontinuous
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)not applicable
Pulse repetition frequency (Hz)not applicable
Duty cycle (%)not applicable
Sonication duration (s)0.16✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)50, 500swept✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatederating
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Single-pulse USNS (160-ms tone bursts) was delivered over left M1 at peak negative pressures stepped from 50 to 500 kPa in 50-kPa increments. In a first subset (n=20), 10 pulses 10 s apart were delivered per pressure step via a threshold-hunting algorithm (electrode-free motor response scoring). In a second subset (n=10, EMG), pressures from 350-500 kPa were tested; 400 kPa (160-msec pulse) gave the best combination of motor success, MEP amplitude, reliability and focality and was adopted for subsequent infralimbic-cortex USNS.

Exposure 2: acute and chronic repeated USNS of the infralimbic cortex (IL)

Target: subgenual anterior cingulate cortex — “infralimbic cortex (IL), the sgACC equivalent in rodents
Device: IGT / Imasonic · Imasonic

Pulse timing
Waveformcontinuous
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)not applicable
Pulse repetition frequency (Hz)not applicable
Duty cycle (%)not applicable
Sonication duration (s)0.16✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)400✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatederating
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Acute USNS was a 10-min session of 60 pulses (each pulse 400 kPa, 160-msec) repeated at a rate of 0.1 Hz over the infralimbic cortex under 1% isoflurane. For chronic USNS, one such session was applied every day for 5 consecutive days (day 29-33), totalling 300 stimuli over 60 min; a Sham group received the same procedure with the transducer deactivated.

Flags from extraction

  • n_subjectsPaper gives group sizes for separate sub-experiments (M1 parameter-finding: 20 electrode-free + 10 EMG; acute USNS: 4; chronic USNS: 15) rather than one exposed total; listed rather than summed.
  • n_sessions_per_subjectVaries by sub-experiment: single session for M1 parameter-finding and acute USNS, 5 daily sessions for chronic USNS.
  • exposures[0].in_situ.pressure_kpaPaper states a skull attenuation coefficient (6.32±2.18% at 500 kHz) used to derate pressure inside the brain but does not give the resulting derated kPa value for specific stimulation levels.
  • exposures[1].target.termsPaper equates the infralimbic cortex (IL) with the rodent sgACC counterpart; target term chosen on that explicit equivalence rather than the vmPFC/prelimbic hierarchy.