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Pulsed focused ultrasound changes nerve conduction of earthworm giant axonal fibers

Sarah H. Yoo, Phillip Croce, Ryan W. Margolin, Stephanie D. Lee, Wonhye Lee

NeuroReport 2017, 28, 229-233 · 10.1097/wnr.0000000000000745

invertebratehealthyinvasive electrophysiology

Abstract

This study examined the effects of pulsed focused ultrasound (FUS) in disrupting nerve conduction. FUS operating at a 210 kHz fundamental frequency was administered to the medial and lateral giant axonal nerve fibers of earthworms in a burst of pulses (1 ms tone burst duration, 20 Hz pulse repetition frequency). The magnitude and latencies of the nerve potentials induced by electrical stimulation were measured under three experimental conditions - (I) no sonication, (II) sonication at 600 mW/cm spatial-peak temporal-average intensity (Ispta), and (III) sonication at 200 mW/cm Ispta. The sonication at 600 mW/cm temporarily decreased the magnitude of the action potential peak (~16%), whereas the baseline peak level was quickly restored in postsonication sessions. Sonication administered at a lower intensity (i.e. 200 mW/cm) did not alter the peak magnitude. The sonication did not alter the nerve conduction velocity. The acoustic intensities used in the experiment did not increase the temperature of the sonicated tissue. The results indicate that axonal neurotransmission can be disrupted temporarily by the application of pulsed FUS, suggesting its potential utility in modulating the functional connectivity established by white matter tracts in the brain.

Abstract via europepmc.

Speciesearthworm (Lumbricus terrestris)
Subjects53 animals
Sessions per subject1
Randomisednot reported
Blindingnot reported
Sham / controlundescribed
Auditory controlnot reported
Readout timingboth
Anaesthesiaanaesthetised
Readoutsinvasive electrophysiologyElectrically-evoked compound nerve action potential magnitude and latency for the median and lateral giant axonal fibers (MGF, LGF), recorded via needle electrodes
Direction of effectinhibitorySonication at 600 mW/cm2 Ispta transiently reduced MGF and LGF action potential magnitude by ~16% without affecting nerve conduction velocity; sonication at 200 mW/cm2 Ispta had no effect on action potential magnitude.
Adverse eventsnone observedThermal measurements at the sonication focus showed no temperature increase across all experimental conditions, indicating the observed effects on nerve conduction were not caused by heating.

Exposures

Exposure 1: FUS to earthworm body (lateral and median giant axonal fibers)

Target: earthworm neurons — “lateral and median giant axonal fibers (LGF/MGF) of the earthworm
Device: Ultran · The Ultran Group Ltd

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)210✓✓
Pulse duration (ms)1✓✓
Pulse repetition frequency (Hz)20✓✓
Duty cycle (%)2pulse duration × PRF gives 2%✓✓
Sonication duration (s)60✓✓
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)945✓✓
Isppa, domain unspecified (W/cm²)10, 30swept✓✓
Ispta, domain unspecified (W/cm²)0.2, 0.6swept✓✓
Protocol, in the paper’s words

Sonication was administered over a tone burst duration of 1 ms at a pulse repetition frequency of 20 Hz (2% duty cycle), delivered continuously for 1 min (sessions f1-f4) at either 600 mW/cm2 or 200 mW/cm2 Ispta, following 5 baseline sessions (B1-B5) and preceding 5 post-sonication sessions (P1-P5); a control group received no sonication.

Flags from extraction

  • n_subjects53 worms were enrolled but 11 were excluded for technical problems; 42 worms (14 per condition) were analysed. n_subjects is recorded as the enrolled total (53) per the enrolled-vs-analysed rule.
  • exposures[0].unspecified_domain.pressure_kpaThe 945 kPa rarefactional pressure is stated only for the 600 mW/cm2 Ispta condition; no corresponding pressure is given for the 200 mW/cm2 condition.
  • exposures[0].unspecified_domain.isppa_w_cm2The paper does not state whether the reported intensities were calibrated in free water or measured/estimated at the worm body; recorded as unspecified domain.
  • sham_typeCondition III is described only as 'without providing actual sonication (i.e. control condition)'; whether the transducer/setup was otherwise identical (powered off) is not explicitly described, so sham mechanism is coded as other.