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Direct Activation of Cortical Neurons in the Primary Somatosensory Cortex of the Rat in Vivo Using Focused Ultrasound

Kush Tripathi, Tongsheng Zhang, Nathan McDannold, Yong-Zhi Zhang, Gösta Ehnholm, Yoshio Okada

Ultrasound in Medicine & Biology 2020 · 10.1016/j.ultrasmedbio.2020.06.003

rodenthealthyinvasive electrophysiology

Abstract

We address the recent controversy over whether focused ultrasound (FUS) activates cortical neurons directly or indirectly by initially activating auditory pathways. We obtained two types of evidence that FUS can directly activate cortical neurons. The depth profile of the local field potential (LFP) in the barrel cortex of the rat in vivo indicated a generator was located within the cortical gray matter. The onset and peak latencies of the initial component p1 were 3.2 ± 0.25 ms (mean ± standard error of the mean) and 7.6 ± 0.12 ms, respectively, for the direct cortical response (DCR), 6.8 ± 0.40 and 14.3 ± 0.54 ms for the FUS-evoked LFP (4 MHz, 3.2 MPa, 50 or 300 µs/pulse, 1-20 pulses at 1 kHz) and 6.9 ± 0.51 and 15.8 ± 0.94 ms for the LFP evoked by 1-ms deflection of the C2 whisker projecting to the same area. The peak latency of the FUS p1 was statistically (t-test) longer than the DCR, but shorter than the whisker p1 at p < 0.005.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects21 animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlnone
Auditory controlnot reported
Readout timingonline
Anaesthesiaanaesthetised
Readoutsinvasive electrophysiologylocal field potential (LFP) recording from barrel cortex via silver-ball/tungsten electrode; laminar depth profiling
Direction of effectexcitatoryFUS evoked a positive-onset LFP (p1) in the barrel column with onset/peak latency between the direct cortical response (DCR) and the whisker-evoked response, and the LFP reversed polarity across cortical depth, indicating direct excitatory activation of cortical neurons within the gray matter.
Adverse eventsnot reported

Exposures

Exposure 1: FUS to barrel column C2 of primary somatosensory cortex (skull removed)

Target: primary somatosensory cortex — “barrel column C2 (S1 barrel cortex)
Device: custom-built

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)4,000✓✓
Pulse duration (ms)0.05, 0.3swept✓✓
Pulse repetition frequency (Hz)1,000✓✓
Duty cycle (%)not reported
Sonication duration (s)0.00000125, 0.02swept✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)3,300✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatederatingsingle value
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)355✓✓
In-situ Ispta (W/cm²)0.00015, 0.036, 0.714swept✓✓
Protocol, in the paper’s words

It produced a train of FUS pulses (each pulse varied between 1.25 and 300 ms depending on the experiment, containing multiple cycles of sinusoidally varying 4-MHz ultrasound). The pulse duration was 50 or 300 ms in most of the experiments. The number of pulses ranged between 1 and 20 at a pulse repetition frequency (PRF) of 1 kHz. Elsewhere the paper labels the shortest pulse duration in microseconds (e.g. '1.25-μs pulse'), so it is unclear whether pulse durations are in ms or μs (see flag).

Flags from extraction

  • exposures[0].timing.pulse_duration_msPulse duration is stated inconsistently: 'The pulse duration was 50 or 300 ms in most of the experiments' and the Figure 3 caption states '1.25 300 ms', yet elsewhere the same intensity calculation labels the shortest duration '1.25-μs pulse', and a 300 ms pulse is physically impossible at the stated 1 kHz PRF (1 ms period). Left not_reported because the correct unit (ms vs μs) cannot be resolved from the text.
  • exposures[0].in_situ.isppa_w_cm2The paper labels 355 W/cm2 as a 'derated' Isptp value 'at the location of interest for measurement (0-3 mm below the cortex)', but this is numerically identical to the intensity value obtained from beam measurements with the lens in place (see free_field discussion elsewhere in Methods), raising doubt about whether true tissue derating was performed or whether this is simply the same water-measured value re-labelled.
  • exposures[0].in_situ.ispta_w_cm2The three listed Ispta values are tied to pulse-duration labels that mix ms and μs inconsistently in the source text (see pulse_duration_ms flag); recorded as stated with unresolved unit ambiguity.
  • n_subjectsThe paper gives the total number of animals used across all experiments in the study (whisker mapping, DCR, and FUS comparisons combined), not a count restricted to animals that received FUS specifically.