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Mechanism of low-frequency, low-intensity ultrasound modulation of the mouse retina

Shun-Yi Zhuo, Hai-Qing Gong, Guo-Feng Li, Wei-Bao Qiu, Hai-Rong Zheng, Pei-Ji Liang

Journal of Neural Engineering 2023 · 10.1088/1741-2552/acd7a4

ex vivo tissuehealthyinvasive electrophysiology

Abstract

Objective. Ultrasound has been shown to modulate the activity of retinal ganglion cells (RGCs) in mice, but the mechanism remains poorly understood. This study aims to address this question. Approach. Multi-electrode recordings together with pharmacological methods were used to investigate the possible cellular/circuitry mechanism(s) underlying the neuronal modulation induced by low-frequency (1 MHz), low-intensity ( I SPTA 0.5 W cm -2 ) ultrasound stimulation. Main results. We found that ultrasound activated mechanosensitive channels (transient receptor potential vanilloid 4 (TRPV4) channels are involved) in Müller cells, causing the release of glutamate, which acts on the extrasynaptic N -methyl-D-aspartate receptors of RGCs, thus leading to the modulation of neuronal activity. Significance. Our results reveal a novel mechanism of low-frequency, low-intensity ultrasound modulation, involving TRPV4 as a mechanosensitive target for ultrasound and glutamate as an essential mediator of neuron-glia communication. These findings also demonstrate that the mechanical-force-mediated pathway is important for retinal signal modulation during visual processes, such as visual accommodation.

Abstract via europepmc.

SpeciesC57BL/6 mouse (isolated retina)
Subjectsnot reported preparations
Sessions per subjectnot applicable
Randomisednot applicable
Blindingnot applicable
Sham / controlnone
Auditory controlnot reported
Readout timingonline
Anaesthesianot applicable
Readoutsinvasive electrophysiologymulti-electrode array recording of retinal ganglion cell spiking
Direction of effectexcitatoryBackground ultrasound increased spontaneous firing (spike count) of ON-RGCs and increased peak firing rate / shortened first-spike latency of light-evoked responses; effect required NMDARs, extrasynaptic NMDARs, and TRPV4.
Adverse eventsnot applicable

Exposures

Exposure 1: background ultrasound stimulation of isolated mouse retina

Target: retina — “retina
Device: Olympus / Panametrics · Olympus NDT, USA · A314S-SU-F1.001N-PTF

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)1,000✓✓
Pulse repetition frequency (Hz)0.333✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 33.3%
Sonication duration (s)1✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)216✓✓
Free-field Isppa (W/cm²)1.5✓✓
Free-field Ispta (W/cm²)0.5✓✓
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

Ultrasound was repeatedly applied to the retina with 1 s duration separated by 2 s intervals 120 times (360 s in total).

Flags from extraction

  • n_subjectsPaper reports retinas-per-experiment (3, 2, 3, 2, 3 retinas across the five pharmacology sub-experiments) but never a single total; cannot determine whether these are overlapping or distinct animals.
  • exposures[0].timing.pulse_repetition_frequency_hzPRF (0.333 Hz) derived from the stated 1 s ON / 2 s OFF period (3 s cycle), a unit conversion from a stated period rather than independent report.
  • exposures[0].timing.sonication_duration_sPaper gives both a 1 s per-pulse ON duration and a 360 s total exposure duration; recorded the 1 s uninterrupted pulse as sonication_duration_s and put the 360 s total in protocol_description, but this is a judgement call under the burst rule.
  • exposures[0].timing.duty_cycle_pctNot directly stated by the paper; not computed from the 1 s ON / 2 s OFF pattern per the rule against deriving duty cycle.
  • exposures[0].free_fieldPressure/intensity measured at the collimator outlet immersed in Ringer's solution (no skull); treated as free-field since paper does not use in-situ/derating language.