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Low-intensity ultrasound suppresses low-Mg 2+ -induced epileptiform discharges in juvenile mouse hippocampal slices

Zhen Zhang, Weibao Qiu, Haiqing Gong, Guofeng Li, Qiuju Jiang, Peiji Liang, Hairong Zheng, Puming Zhang

Journal of Neural Engineering 2019, 16, 036006 · 10.1088/1741-2552/ab0b9a

ex vivo tissueepilepsyinvasive electrophysiology

Abstract

Objective It has been shown that low-intensity ultrasound (LIUS) can suppress seizures in some laboratory studies. However, the mechanism of the suppression effect of LIUS remains unclear. The goal of this study is to investigate the modulation effects of focused LIUS on epileptiform discharges in mouse hippocampal slices as well as the underlying mechanism. Approach Epileptiform discharges in hippocampal slices of 8 d-old mice were induced by low-Mg 2+ artificial cerebrospinal fluid and recorded by a micro-electrode array in vitro. LIUS was delivered to hippocampal slices to investigate its modulation effects on epileptiform discharges. Pharmacological experiments were conducted to study the mechanism of the modulation effects. Main results LIUS suppressed the amplitude, rate and duration of ictal discharges. For inter-ictal discharges, LIUS suppressed the amplitude but facilitated the rate. LIUS suppressed the spontaneous spiking activities of pyramidal neurons in CA3, and the suppression effect was eliminated by Kaliotoxin. The suppression effect of LIUS on epileptiform discharges was weakened when the perfusion was mixed with Kaliotoxin. Significance Those findings demonstrate that LIUS suppresses the epileptiform discharges in 8 d-old mouse hippocampal slices and that its suppression effect can mainly attributed to the activation of mechanosensitive Kv1.1 channels.

Abstract via europepmc.

SpeciesC57BL/6 mouse (8-day-old)
Subjectsnot reported preparations
Sessions per subjectnot applicable
Randomisednot applicable
Blindingnot applicable
Sham / controlnone
Auditory controlnot reported
Readout timingboth
Anaesthesianot applicable
Readoutsinvasive electrophysiologymicroelectrode array (MEA) local field potential and single-unit spike recordings; epileptiform discharge rate/duration/amplitude analysis
Direction of effectinhibitoryLIUS suppressed the rate, amplitude and duration of ictal discharges (IDs) and suppressed the amplitude (but facilitated the rate) of inter-ictal discharges (IIDs); it also suppressed spontaneous spiking of CA3 pyramidal neurons. This suppression was attenuated by the Kv1.1 blocker kaliotoxin.
Adverse eventsnot applicable

Exposures

Exposure 1: Focused low-intensity ultrasound (LIUS) applied to hippocampal slice

Target: hippocampal formation — “hippocampal slice (CA3, CA1, dentate gyrus, and subiculum)
Device: Olympus / Panametrics · Olympus NDT · A314S

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)0.4✓✓
Pulse repetition frequency (Hz)2,000✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 80%
Sonication duration (s)1✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)119✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)0.0078, 0.0709, 0.126swept✓✓
In-situ estimatenot applicable
In-situ pressure (kPa)not applicable
In-situ Isppa (W/cm²)not applicable
In-situ Ispta (W/cm²)not applicable
Protocol, in the paper’s words

Sonication parameters: sonication duration (SD) = 1 s, inter-stimulus interval (ISI) = 3 s, tone burst duration (TBD) = 0.4 ms, pulse repetition period (PRP) = 0.5 ms, repeated for a total LIUS administration period of 900 s (15 min) per slice.

Flags from extraction

  • n_subjectsDifferent sub-analyses report different slice counts (e.g. n=10 for main suppression statistics, n=18 for CA3 VPT analysis, n=5 for KTX pharmacology); no single total number of slices/mice used in the whole study is stated.
  • exposures[0].timing.pulse_repetition_frequency_hzPaper states pulse repetition PERIOD (PRP=0.5 ms) rather than PRF directly; converted via PRF=1/PRP (unit conversion of a stated period, per instructions).
  • exposures[0].in_situ.pressure_kpaValues described as 'in hippocampal slice' were derived from a hydrophone scan of the free field mapped onto the expected slice position, not a measurement through/within intact tissue; classified as in_situ based on the paper's own wording ('in hippocampal slice') but flagged for domain ambiguity.