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Ketamine Inhibits Ultrasound Stimulation-Induced Neuromodulation by Blocking Cortical Neuron Activity

Sungmin Han, Minkyung Kim, Hyungmin Kim, Hyunjoon Shin, Inchan Youn

Ultrasound in Medicine & Biology 2018 · 10.1016/j.ultrasmedbio.2017.11.008

rodentin vitro cellhealthybehaviourcellular imaginghistology molecular

Abstract

Ultrasound (US) can be used to noninvasively stimulate brain activity. However, reproducible motor responses evoked by US are only elicited when the animal is in a light state of anesthesia. The present study investigated the effects of ketamine on US-induced motor responses and cortical neuronal activity. US was applied to the motor cortex of mice, and motor responses were evaluated based on robustness scores. Cortical neuronal activity was observed by fluorescence calcium imaging. US-induced motor responses were inhibited more than 20 min after ketamine injection, and US-triggered Ca 2+ transients in cortical neurons were effectively blocked by ketamine. Our results indicate that ketamine suppresses US-triggered Ca 2+ transients in cortical neurons and, therefore, inhibits US-induced motor responses in a deep anesthetic state.

Abstract via europepmc.

Speciesmouse (C57BL/6)
Subjects8, 6, 17, 3swept animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlno treatment control
Auditory controlnot reported
Readout timingboth
Anaesthesiaanaesthetised
Readoutsbehaviour, cellular imaging, histology moleculartail-flick motor response robustness scoring; fluorescence calcium imaging (OGB-1 AM) of cultured cortical neurons; H&E staining
Direction of effectexcitatoryUltrasound evoked motor responses and triggered Ca2+ transients in cortical neurons (excitatory activation); ketamine dose-dependently blocked the US-evoked Ca2+ transients and suppressed motor responses in a deep anesthetic state.
Adverse eventsnone observedObvious morphologic changes or damage was not induced by US. Additionally, tissue bleeding or cytoplasmic swelling was not found in the stimulated cortical regions compared with control brain tissue.

Exposures

Exposure 1: Motor cortex sonication (mice, in vivo)

Target: primary motor cortex — “motor cortex
Device: other named manufacturer · EofE Ultrasonics Co., Ltd

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)350✓✓
Pulse duration (ms)0.23✓✓
Pulse repetition frequency (Hz)1,500✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 34.5%
Sonication duration (s)0.0667✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)60, 690swept✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)1.16, 13.55swept✓✓
In-situ estimatenot reported
In-situ pressure (kPa)not reported
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Two function generators were serially connected to generate bursts of pulsed waves, where the first function generator regulated the PRF, the SD and the inter-stimulus interval (ISI). Sonication was induced with 350-kHz FF, 1.5-kHz PRF, 0.23-ms TBD, and 66.67-ms SD; the robustness scores were evaluated for AIs varied 1.16-13.55 W/cm2 (0.06-0.69 MPa), and a fixed AI of 3.46 W/cm2 was used for the onset-time and histologic safety experiments.

Exposure 2: Cultured cortical neurons (in vitro)

Target: cultured neurons — “cultured cortical neurons
Device: other named manufacturer · EofE Ultrasonics Co., Ltd

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)350✓✓
Pulse duration (ms)0.23✓✓
Pulse repetition frequency (Hz)1,500✓✓
Duty cycle (%)not reportedpulse duration × PRF gives 34.5%
Sonication duration (s)0.0667✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)0.056✓✓
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 was induced with the same parameters described earlier (350-kHz FF, 1.5-kHz PRF, 0.23-ms TBD, 66.67-ms SD) and the AI was set to 56 mW/cm2, measured in the degassed water; fluorescence was recorded for 3 min with US applied 1 min after recording started.

Flags from extraction

  • n_subjectsPaper reports separate mouse cohorts for different sub-experiments (8, 6, 17, and 3 mice) without stating an overall total exposed to ultrasound; unclear whether cohorts overlap.
  • exposures[0].free_field.isppa_w_cm2The 1.16-13.55 W/cm2 range (robustness-score sweep) and the fixed 3.46 W/cm2 value (used for onset-time and histology experiments) are combined in one list; 3.46 W/cm2 has no separately stated pressure equivalent.
  • exposures[*].free_fieldDomain assigned as free_field because the Methods state AI/pressure values were measured via hydrophone in a degassed water tank (and the in vitro AI is explicitly stated as measured in degassed water); the paper does not label every individual number with a domain.
  • exposures[0].timing.duty_cycle_pctNot stated as a percentage in the text; not computed from pulse duration and PRF per instructions.