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Inhibitory effect of ultrasonic stimulation on the voltage-dependent potassium currents in rat hippocampal CA1 neurons

Kun Cui, Shuai Zhang, Jinyao Sun, Xueying Zhang, Chong Ding, Guizhi Xu

BMC Neuroscience 2019, 20 · 10.1186/s12868-018-0485-1

ex vivo tissuehealthyinvasive electrophysiology

Abstract

Background Transcranial ultrasonic stimulation is a novel noninvasive tool for neuromodulation, and has high spatial resolution and deep penetration. Although it can increase excitation of neurons, its effects on neuron are poorly understood. This study was to evaluate effect of ultrasonic stimulation (US) on neurons in vitro. In this paper, the effect of US on the excitability and voltage-dependent [Formula: see text] currents of CA1 pyramidal neurons in the rat hippocampus was studied using patch clamp. Results Our results suggest that US increased the spontaneous firing rate and inhibited transient outward potassium current ([Formula: see text]) and delayed rectifier potassium current ([Formula: see text]. Furthermore, US altered the activation of [Formula: see text] channels, inactivation and recovery properties of [Formula: see text] channels. After US, the [Formula: see text] activation curves significantly moved to the negative voltage direction and increased its slope factor. Moreover, the data showed that US moved the inactivation curve of [Formula: see text] to the negative voltage and increased the slope factor. Besides, US delayed the recovery of [Formula: see text] channel. Conclusions Our data indicate that US can increase excitation of neurons by inhibiting potassium currents. Different US decreased the voltage sensitivity of [Formula: see text] activation differentially. Moreover, the more time is needed for US to make the [Formula: see text] channels open again after inactivating. US may play a physiological role by inhibiting voltage-dependent potassium currents in neuromodulation. Our research can provide a theoretical basis for the future clinical application of ultrasound in neuromodulation.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects12 preparations
Sessions per subjectnot applicable
Randomisednot applicable
Blindingnot applicable
Sham / controlundescribed
Auditory controlnot reported
Readout timingnot reported
Anaesthesianot applicable
Readoutsinvasive electrophysiologywhole-cell patch clamp recordings of spontaneous action potentials and voltage-dependent potassium currents (IA, IK) in CA1 pyramidal neurons
Direction of effectexcitatoryUS increased spontaneous firing rate and action potential amplitude while inhibiting transient outward (IA) and delayed rectifier (IK) potassium currents, shifting their voltage dependence and slowing IA recovery from inactivation.
Adverse eventsnot applicable

Exposures

Exposure 1: Ultrasonic stimulation of hippocampal CA1 brain slices

Target: CA1 — “rat hippocampal CA1 neurons
Device: Olympus / Panametrics · Olympus · V308

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)20✓✓
Pulse repetition frequency (Hz)20✓✓
Duty cycle (%)50pulse duration × PRF gives 40%, which disagrees with the stated value✓✓
Sonication duration (s)900✓✓
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 applicable
In-situ pressure (kPa)not applicable
In-situ Isppa (W/cm²)not applicable
In-situ Ispta (W/cm²)not applicable
Isppa, domain unspecified (W/cm²)0.015, 0.03swept✓✓
Protocol, in the paper’s words

The ultrasound settings were 0.5 MHz center frequency, 20 Hz pulse repetition frequency, 50% duty cycle and 20 ms pulse length, and the pulse-average ultrasound intensities were 15 mW/cm2 or 30 mW/cm2. All stimulations lasted 15 min. The transducer (diameter 24 mm) was immersed in aCSF 1.5 cm above the brain slice and stimulated the entire slice.

Flags from extraction

  • sham_typeControl condition was brain slices with no stimulation applied; the paper does not describe whether an inactive transducer was left in place, so the sham mechanism is not fully described.
  • exposures[0].timing.pulse_duration_msStated pulse length (20 ms) does not exactly match duty_cycle/PRF (50%/20 Hz = 25 ms); recorded as stated, not corrected.
  • exposures[0].unspecified_domain.isppa_w_cm2Paper reports 'pulse-average ultrasound intensity' without stating whether this was measured in water/free field or at the slice; placed in unspecified_domain.
  • readout_timingPaper does not state whether patch-clamp recordings were made during or after the 15-minute ultrasound exposure.