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Influence of behavioral state on the neuromodulatory effect of low-intensity transcranial ultrasound stimulation on hippocampal CA1 in mouse

Xingran Wang, Yiyao Zhang, Kaiqing Zhang, Yi Yuan

NeuroImage 2021, 241, 118441 · 10.1016/j.neuroimage.2021.118441

rodenthealthyinvasive electrophysiology

Abstract

In process of brain stimulation, the influence of any external stimulus depends on the features of the stimulus and the initial state of the brain. Understanding the state-dependence of brain stimulation is very important. However, it remains unclear whether neural activity induced by ultrasound stimulation is modulated by the behavioral state. We used low-intensity focused ultrasound to stimulate the hippocampal CA1 regions of mice with different behavioral states (anesthesia, awake, and running) and recorded the neural activity in the target area before and after stimulation. We found the following: (1) there were different spike firing rates and response delays computed as the time to reach peak for all behavioral states; (2) the behavioral state significantly modulates the spike firing rate linearly increased with an increase in ultrasound intensity under different behavioral states; (3) the mean power of local field potential induced by TUS significantly increased under anesthesia and awake states; (4) ultrasound stimulation enhanced phase-locking between spike and ripple oscillation under anesthesia state. These results suggest that ultrasound stimulation-induced neural activity is modulated by the behavioral state. Our study has great potential benefits for the application of ultrasound stimulation in neuroscience.

Abstract via europepmc.

Speciesmouse (C57BL/6, male)
Subjects78 animals
Sessions per subjectnot reported
Randomisedyes
Blindingnot reported
Sham / controlnone
Auditory controldeafened subjects
Readout timingboth
Anaesthesiaboth
Readoutsinvasive electrophysiologyExtracellular single/multi-unit spike recording and local field potential (LFP) recording in hippocampal CA1
Direction of effectexcitatoryTUS increased spike firing rate and LFP power in hippocampal CA1 across anesthesia, awake, and running states, with the magnitude of the increase strongly dependent on behavioral state (largest under anesthesia, smallest during running); firing rate increased linearly with ultrasound intensity in all states.
Adverse eventsnot reported

Exposures

Exposure 1: TUS to hippocampal CA1 under anesthesia, awake, and running behavioral states (Experiments A-D)

Target: CA1 — “hippocampal CA1
Device: Olympus / Panametrics · Olympus · V323-SU (focused ultrasound transducer)

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)2,250✓✓
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 estimatemeasurementsingle value
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Pressure, domain unspecified (kPa)150, 300, 450, 600, 750swept✓✓
Isppa, domain unspecified (W/cm²)0.75, 3, 6.75, 12, 18.75swept✓✓
Protocol, in the paper’s words

16 trials were averaged per animal. Ultrasound intensity was fixed at 0.45 MPa for Experiments A (single-parameter neural activity, N=27 across 3 behavioral-state groups) and C (sleep, N=3); Experiment B (N=30) swept intensity across 0.15, 0.3, 0.45, 0.6 and 0.75 MPa to relate firing rate to intensity across states; Experiment D (N=18) repeated the single-parameter stimulation in chemically deafened mice to test for auditory confounds. The interval between TUS deliveries was 10 s within each recording session.

Flags from extraction

  • exposures[0].timing.pulse_duration_msOnly 'stimulation duration' (400 ms, the train length), PRF (1 kHz) and duty cycle (50%) are stated; individual pulse width within the train is not stated in words and was not computed from duty cycle/PRF.
  • exposures[0].in_situ.pressure_kpaThe five intensity levels (0.15-0.75 MPa) were measured 'under the skull' with a needle hydrophone per the methods (in situ, via direct measurement), and are listed together as a single exposure per the instruction to record within-target parameter sweeps as lists rather than separate exposures.
  • n_sessions_per_subjectPaper describes 16 trials averaged per experiment but does not state how many separate sessions each individual mouse underwent.