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The Effect of Low-Intensity Transcranial Ultrasound Stimulation on Neural Oscillation and Hemodynamics in the Mouse Visual Cortex Depends on Anesthesia Level and Ultrasound Intensity

Yi Yuan, Kaiqing Zhang, Yiyao Zhang, Jiaqing Yan, Zhijie Wang, Xingrang Wang, Mengyang Liu, Xiaoli Li

IEEE Transactions on Biomedical Engineering 2021, 68, 1619-1626 · 10.1109/tbme.2021.3050797

rodenthealthyinvasive electrophysiologycerebral haemodynamics

Abstract

Objective Low-intensity transcranial ultrasound stimulation (TUS) can induce motor responses, neural oscillation and hemodynamic responses. Early studies demonstrated that the motor responses evoked by TUS critically depend on anesthesia levels and ultrasound intensity. However, the neural mechanism of how anesthesia levels and ultrasound intensity influence on brain responses during TUS has never been explored yet. To investigate this question, we applied different anesthesia levels and ultrasound intensities on the visual cortex of mouse and observed neural oscillation change and hemodynamic responses during TUS. Methods low-intensity ultrasound was delivered to mouse visual cortex under different anesthesia levels, and simultaneous recordings for local field potentials (LFPs) and hemodynamic responses were carried out to measure and analyze the changes quantitatively. Results (i) The change of mean amplitude and mean relative power of sharp wave-ripple (SPW-R) in LFPs induced by TUS decreased as the anesthesia level increased (from awake to 1.5% isoflurane). (ii) The hemodynamic response level induced by TUS decreased as the anesthesia level increased (from awake to1.5% isoflurane). (iii) The coupling strength between neural activities and hemodynamic responses was dependent on anesthesia level. (iv) The neural activities and hemodynamic responses increase as a function of ultrasound intensity. Conclusion These results support that the neural activities and hemodynamic response of the mouse visual cortex induced by TUS are related to the anesthesia level and ultrasound intensity. Significance This finding suggests that careful maintenance of anesthesia level and ultrasound intensity is required to acquire accurate LFP and hemodynamic data from samples with TUS.

Abstract via europepmc.

Speciesmouse (C57BL)
Subjects42 animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlnone
Auditory controlnot reported
Readout timingboth
Anaesthesiaboth
Readoutsinvasive electrophysiology, cerebral haemodynamicsLocal field potential recording (silver needle electrode) for sharp wave-ripple (SPW-R) detection; intrinsic signal optical imaging of deoxyhemoglobin (HbR) hemodynamic responses.
Direction of effectexcitatoryTUS induced sharp wave-ripple (SPW-R) neural activity and hemodynamic (HbR) responses that decreased in amplitude with increasing anesthesia depth (awake > 0.3% > 0.9% > absent at 1.5% isoflurane) and increased with increasing ultrasound intensity.
Adverse eventsnot reportedIn our experiment, the maximum ultrasound intensity I is $2 7 ~ \mathrm { W / c m ^ { 2 } }$ and pulse duration t is 0.4s. We can get that the temperature rise is ${ \sim } 0 . 1 2 ^ { \circ } \mathrm { C }$ , which is lower than the damage threshold.

Exposures

Exposure 1: TUS of visual cortex across anesthesia levels and ultrasound intensities

Target: visual cortex — “mouse visual cortex (AP=-3.8, ML=2.5)
Device: not reported

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)not reportedimplied by duty cycle ÷ PRF: 0.5 ms (not stated by the paper)
Pulse repetition frequency (Hz)1,000✓✓
Duty cycle (%)50✓✓
Sonication duration (s)0.4✓✓
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)50, 100, 300, 500, 550, 700, 900swept✓✓
Isppa, domain unspecified (W/cm²)0.08, 0.3, 3, 8.3, 10.1, 16.3, 27swept✓✓
Protocol, in the paper’s words

A circular skull section (3.5 mm diameter) was removed to expose the visual cortex directly for TUS delivery. Mice were tested under 0% (awake), 0.3%, 0.9% and 1.5% isoflurane using a fixed maximum ultrasound pressure of 0.55 MPa (10.1 W/cm2 Isppa); a separate group of mice was tested at 0.3% and 0.9% isoflurane across a sweep of ultrasound pressures (0.05-0.9 MPa). Sixteen trials were averaged per condition; LFP and hemodynamic responses were quantified over a 0-3 s window from TUS onset.

Flags from extraction

  • exposures[0].timing.pulse_duration_msThe 400 ms value is explicitly labelled 'stimulation duration' (SD, recorded as sonication_duration_s) in Methods, but the Discussion separately refers to '0.4s' as 'pulse duration' in a temperature calculation; duty cycle (50%) and PRF (1000 Hz) would imply an internal tone-burst of 0.5 ms, but this is not stated directly and is not computed here.
  • exposures[0].unspecified_domainPaper does not state whether pressures/intensities are free-field or in-situ; the visual cortex was directly exposed via craniotomy (no skull in the acoustic path), but this is not discussed by the authors as a domain distinction.
  • exposures[0].unspecified_domain.pressure_kpaList combines the single fixed pressure used in the anesthesia-level experiment (550 kPa) with the separate pressure sweep used in the ultrasound-intensity experiment (50-900 kPa); these are two different sub-experiments, not one continuous sweep.
  • deviceNo transducer manufacturer or model is given in the main text; setup is described as similar to a previous study by the same group.