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Microscopic deconstruction of cortical circuit stimulation by transcranial ultrasound

Théo Lemaire, Yi Yuan, Celia Gellman, Amy M. LeMessurier, Sarah R. Haiken Dray, Justin P. Little, Robert C. Froemke, Shy Shoham

2024 · 10.1101/2024.10.10.617091

rodenthealthycellular imaging

Abstract

Transcranial Ultrasound Stimulation (TUS) can noninvasively and reversibly perturb neuronal activity, but the mechanisms by which ultrasound engages brain circuits to induce functional effects remain unclear. To elucidate these interactions, we applied TUS to the cortex of awake mice and concurrently monitored local neural activity at the acoustic focus with two-photon calcium imaging. We show that TUS evokes highly focal responses in three canonical neuronal populations, with cell-type-specific dose dependencies. Through independent parametric variations, we demonstrate that evoked responses collectively scale with the time-average intensity of the stimulus. Finally, using computational unmixing we propose a physiologically realistic cortical circuit model that predicts TUS-evoked responses as a result of both direct effects and local network interactions. Our results provide a first direct evidence of TUS’s focal effects on cortical activity and shed light on the complex circuit mechanisms underlying these effects, paving the way for TUS’s deployment in clinical settings.

Abstract via europepmc.

Speciesmouse
Subjects6, 10, 7swept animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlinactive transducer, sound only
Auditory controlsound only sham, deafened subjects
Readout timingonline
Anaesthesiaawake
Readoutscellular imagingTwo-photon calcium imaging (GCaMP6s in Thy1+ pyramidal, SST+ and PV+ interneurons; GCaMP7f in auditory cortex) of layer 2/3 cortical neurons
Direction of effectexcitatoryFocused 2.1 MHz TUS evoked focal, cell-type-specific increases in calcium activity in Thy1+, SST+ and PV+ V1 neurons that scaled with time-average acoustic intensity (ISPTA); SST responses declined modestly at the highest intensities, attributed to network-mediated inhibition from recruited PV interneurons rather than a change in direct sign of the effect.
Adverse eventsnot reported

Exposures

Exposure 1: Focused TUS to primary visual cortex (V1), 2.1 MHz, pressure and duty-cycle sweeps

Target: primary visual cortex — “primary visual cortex (V1)
Device: custom-built · Beijing C.C.W. Ultrasonic Science & Technology, China (PZT-4 ceramic ring element, custom-assembled transducer)

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)2,100✓✓
Pulse duration (ms)not reported
Pulse repetition frequency (Hz)100✓✓
Duty cycle (%)5, 80swept✓✓
Sonication duration (s)0.2✓✓
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
Pressure, domain unspecified (kPa)0, 800swept?
Ispta, domain unspecified (W/cm²)0, 15.8swept?
Protocol, in the paper’s words

TUS delivered as 200 ms pulse trains (internal PRF 100 Hz) through a coaxial annular transducer aligned with the 2-photon objective; independent sweeps of peak pressure amplitude (0-0.8 MPa at fixed 50% duty cycle) and duty cycle (5-80% at fixed 0.8 MPa) were performed. For each condition, 16 consecutive stimuli were presented with a 25-28 s inter-trial interval; no more than 15 conditions were explored per session (imaging time <2 hours).

Flags from extraction

  • n_subjectsDifferent sub-experiments (dose-response sweeps, offset control, buzzing control, A1/deafening control) report different, partially overlapping animal counts (e.g. SST n=6, Thy1 n=10, PV n=7, offset n=3, buzzing n=4); no single total number of animals used in the study is stated.
  • exposures[0].timing.pulse_duration_msPaper gives duty cycle (5-80%) and PRF (100 Hz) but never states an explicit pulse duration in ms; computing it from duty cycle/PRF would be arithmetic not stated by the paper, so left not_reported per the burst rule.
  • exposures[0].unspecified_domain.pressure_kpaPeak pressure amplitude (0-0.8 MPa) is given without specifying whether it is a free-field or in-situ/at-target value; placed in unspecified_domain.
  • exposures[0]A second target (primary auditory cortex, A1) was also stimulated with the same transducer/frequency as a confound-control experiment (deafened animal, n=1); this was not modelled as a separate exposure because it served as a mechanistic control rather than a primary parametric target.