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Low-frequency, low-intensity ultrasound modulates light responsiveness of mouse retinal ganglion cells

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

Journal of Neural Engineering 2022 · 10.1088/1741-2552/ac7d75

ex vivo tissuehealthyinvasive electrophysiology

Abstract

Objective . Ultrasound modulates the firing activity of retinal ganglion cells (RGCs), but the effects of lower-frequency, lower-intensity ultrasound on RGCs and underlying mechanism(s) remain poorly understood. This study aims to address these questions. Approach . Multi-electrode recordings were used in this study to record the firing sequences of RGCs in isolated mouse retinas. RGCs' background firing activities as well as their light responses were recorded with or without ultrasound stimulation. Cross-correlation analyses were performed to investigate the possible cellular/circuitry mechanism(s) underlying ultrasound modulation. Main results . It was found that ultrasound stimulation of isolated mouse retina enhanced the background activity of ON-RGCs and OFF-RGCs. In addition, background ultrasound stimulation shortened the light response latency of both ON-RGCs and OFF-RGCs, while enhancing part of the RGCs' (both ON- and OFF-subtypes) light response and decreasing that of the others. In some ON-OFF RGCs, the ON- and OFF-responses of an individual cell were oppositely modulated by the ultrasound stimulation, which suggests that ultrasound stimulation does not necessarily exert its effect directly on RGCs, but rather via its influence on other type(s) of cells. By analyzing the cross-correlation between the firing sequences of RGC pairs, it was found that concerted activity occurred during ultrasound stimulation differed from that occurred during light stimulation, in both spatial and temporal aspects. These results suggest that the cellular circuits involved in ultrasound- and light-induced concerted activities are different and glial cells may be involved in the circuit in response to ultrasound. Significance . These findings demonstrate that ultrasound affects neuronal background activity and light responsiveness, which are critical for visual information processing. These results may also imply a hitherto unrecognized role of glial cell activation in the bidirectional modulation effects of RGCs and may be critical for the nervous system.

Abstract via europepmc.

Speciesmouse (C57BL/6)
Subjects17 preparations
Sessions per subjectnot applicable
Randomisednot applicable
Blindingnot applicable
Sham / controlnone
Auditory controlnot reported
Readout timingonline
Anaesthesianot applicable
Readoutsinvasive electrophysiologyMulti-electrode array (MEA) recording of retinal ganglion cell (RGC) spike firing rate, first-spike latency and cross-correlation histograms, with and without light stimulation
Direction of effectbidirectionalBackground ultrasound increased spontaneous firing of both ON- and OFF-RGCs and shortened light-response latency in all responsive cells, but light-evoked firing rate was increased in some RGCs and decreased in others (including opposite modulation of the ON- and OFF-components within some ON-OFF RGCs), suggesting an indirect, glial-cell-mediated mechanism rather than a uniform excitatory or inhibitory action.
Adverse eventsnot applicable

Exposures

Exposure 1: 1 MHz background ultrasound stimulation of isolated retina

Target: retinal explant — “isolated mouse retina
Device: Olympus / Panametrics · Olympus NDT · A314S-SU-F1.001N-PTF

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)1,000?
Pulse repetition frequency (Hz)not reported
Duty cycle (%)not reported
Sonication duration (s)360?
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

The ultrasound transducer, mounted with a conical acoustic collimator positioned 3 mm above the MEA with the collimator tip immersed in the perfusate, delivered background ultrasound at the retina during 90 light-response trials divided into control, ultrasound and recovery periods; background ultrasound was provided for 360 s at the 31st-60th trials, with the ON-transients of light stimulation synchronised to the ultrasound stimulation.

Flags from extraction

  • exposures[0].free_fieldThe retina was bathed directly in Ringer's solution with the collimator tip immersed in the perfusate (no skull/tissue path); values are placed in free_field per the instruction that for a bath/dish preparation with no skull or tissue path, in_situ is null.
  • exposures[0].timing.pulse_duration_ms,exposures[0].timing.sonication_duration_sThe paper describes a 1-s ultrasound-ON / 2-s ultrasound-OFF repeating pattern for 120 repeats (360 s total background stimulation). This was recorded as pulse_duration_ms=1000 (the 1 s 'ON' period) and sonication_duration_s=360 (the total 360 s background stimulation block); PRF and duty cycle for the ON/OFF pattern itself are not explicitly stated as such by the paper.
  • n_subjects17 retinas were used in total for the reported cell counts (235 ON-RGCs, 58 OFF-RGCs, 97 ON-OFF RGCs); subject_unit recorded as 'preparation' (one isolated retina per animal is assumed but not explicitly stated).