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Transcranial Ultrasound Stimulation Reverses Behavior Changes and the Expression of Calcium-Binding Protein in a Rodent Model of Schizophrenia

Che-Wen Tsai, Shih-Jen Tsai, Yi-Ju Pan, Hsin-Mei Lin, Tsung-Yu Pan, Feng-Yi Yang

Neurotherapeutics 2022, 19, 649-659 · 10.1007/s13311-022-01195-x

rodentschizophreniabehaviourhistology molecular

Abstract

Cognitive dysfunctions are a core feature of schizophrenia that may be linked to abnormalities in gamma-aminobutyric-acid (GABA)ergic neurons. Traditional antipsychotics show poor efficacy in treating cognitive symptoms. The purpose of this study was to investigate the restorative role of transcranial ultrasound stimulation (TUS) in counteracting dizocilpine (MK-801)-induced cognitive deficits and GABAergic interneuron dysfunction in a simulation of schizophrenia. Some rats subjected to MK-801 administration were treated with low-intensity pulsed ultrasound (LIPUS) daily for 5 days, while other rats subjected to MK-801 administration received no LIPUS treatment. After LIPUS treatment, the neuroprotective effects of LIPUS in the LIPUS-treated rats were assessed through behavioral analysis, western blotting, and histological observations. Compared with the MK-801-treated group, the MK-801 plus LIPUS-treated rats revealed a preference for novel objects. The MK-801 plus LIPUS-treated rats also exhibited a significant decrease in swim times compared to the MK-801-treated rats. LIPUS stimulation significantly increased hippocampal levels of CB and PV and restored the cell densities of PV + and CB + in the cingulate cortex in the MK-801 plus LIPUS-treated group. In addition, LIPUS stimulation rebalanced the BDNF levels in the hippocampus and medial prefrontal cortex. Our findings indicate that LIPUS improves cognitive deficits and ameliorates neuropathology in MK-801-treated rats. These results suggest that LIPUS may constitute a potential novel therapeutic approach for the treatment of schizophrenia.

Abstract via europepmc.

Speciesrat (Sprague-Dawley)
Subjects8, 8swept animals
Sessions per subject5
Randomisedyes
Blindingnot reported
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutsbehaviour, histology molecularspontaneous object recognition task; Morris water maze reversal learning; open field locomotor activity; western blot (GAD67, CB, PV, BDNF, GDNF, VEGF); immunofluorescence (PV+ and CB+ cell counts in cingulate cortex)
Direction of effectnot assessedLIPUS reversed MK-801-induced deficits: improved novel-object discrimination and reversal-learning performance, restored hippocampal CB and PV levels, restored PV+/CB+ cell densities in cingulate cortex, and rebalanced BDNF levels in hippocampus and mPFC; these are behavioral-rescue and molecular outcomes rather than a measured direction of neuromodulatory effect.
Adverse eventsnot reported

Exposures

Exposure 1: LIPUS to brain region near bregma in MK-801 rat model of schizophrenia

Target: hippocampus — “desired region (3.0 mm posterior and 2.5 mm lateral to the bregma) of the brain
Device: Olympus / Panametrics · Panametrics · A392S

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)50✓✓
Pulse repetition frequency (Hz)1✓✓
Duty cycle (%)5pulse duration × PRF gives 5%✓✓
Sonication duration (s)300✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)not reported
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)0.528✓✓
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

The focused transducer was positioned using a stereotaxic apparatus in order to direct the acoustic beam to the desired region (3.0 mm posterior and 2.5 mm lateral to the bregma) of the brain. Each rat hemisphere was treated by LIPUS daily with triple sonications. The duration of each sonication was 5 min, and there was an interval of 5 min between each sonication. LIPUS would be turned on when the LIPUS group and the MK-801 + LIPUS group accepted treatment while the Sham group and MK-801 group were just anesthetized.

Flags from extraction

  • n_subjectsPaper reports per-group n=8 for behavioral cohorts (LIPUS group and MK-801+LIPUS group are the two groups receiving ultrasound) but never a combined total; a subset of n=5 per group was used for histology. Recorded as the list of the two ultrasound-exposed group sizes [8,8].
  • n_sessions_per_subjectComputed from 'daily with triple sonications' over the treatment period; paper states LIPUS stimulation lasted 5 days (sacrificed 24 h after last treatment), so 5 daily sessions (each with 3 sonications) is inferred from the described schedule.
  • exposures[0].target.termsTarget coordinates (3.0 mm posterior, 2.5 mm lateral to bregma) are not explicitly named by the paper as a specific brain structure; classified as hippocampus based on typical rat stereotaxic coordinates and the paper's focus on hippocampal CB/PV/BDNF outcomes, but this is an inference and not the paper's own terminology.
  • direction_of_effectStudy reports behavioral-rescue and molecular (PV/CB/BDNF) outcomes rather than a measured direction of neuromodulatory (excitatory/inhibitory) neural effect, so 'not_assessed' was used.
  • adverse_eventsPaper does not include a dedicated safety/histology evaluation section (unlike the companion Song et al. 2022 paper using the same device/parameters), so adverse_events could not be confirmed from the text.