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Effects of Transcranial Ultrasound Stimulation on Blood Oxygen Metabolism and Brain Rhythms in Nitroglycerin-Induced Migraine Mice

Nannan Bian, Yi Yuan, Xiaoli Li

Neuromodulation: Technology at the Neural Interface 2024, 27, 824-834 · 10.1016/j.neurom.2023.12.007

rodentotherbehaviourinvasive electrophysiologyhistology molecularcerebral haemodynamics

Abstract

Objectives In this study, we aimed to investigate the regulatory mechanism of transcranial ultrasound stimulation (TUS) on nitroglycerin-induced migraine in mice. Materials and methods The experiment was divided into four groups, namely, the normal saline control group (n = 9), ultrasound stimulation control group (n = 6), nitroglycerin-induced migraine group (n = 9), and ultrasound stimulation group (n = 9). The behavior, blood oxygen metabolism, and brain rhythm distribution of the four groups were analyzed. Results We found that after TUS, the movement time and speed of mice with migraine are modulated to those of the control groups, and the number of head scratching and grooming events is significantly reduced. TUS increased the deoxygenated hemoglobin, and the power of the 4-to-40 Hz frequency band of local field potentials in the cortex of migraine mice. TUS also decreased the expression of plasma calcitonin gene-related peptide and cortical c-Fos protein. Conclusions Ultrasound stimulation can regulate brain rhythm and blood oxygen metabolism and reduce migraine symptoms in mice. The regulatory mechanism may be related to reducing calcitonin gene-related peptide in blood vessels.

Abstract via europepmc.

Speciesmouse (C57BL/6N)
Subjects6, 9swept animals
Sessions per subjectnot reported
Randomisednot reported
Blindingnot reported
Sham / controlnone
Auditory controlnot reported
Readout timingboth
Anaesthesiaawake
Readoutsbehaviour, invasive electrophysiology, histology molecular, cerebral haemodynamicsOpen-field behavior (movement time/speed, head scratching, grooming); intrinsic signal optical imaging (ISOI) of cortical deoxyhemoglobin (HbR); single-channel LFP power spectra; plasma CGRP ELISA; cortical CGRP and c-Fos Western blot
Direction of effectbidirectionalTUS decreased the NTG-elevated 0.5-4 Hz LFP power band while increasing the NTG-suppressed 4-40 Hz power bands, returning both toward control levels; TUS also kept cortical HbR close to control levels (inhibiting the NTG-induced decrease) and reduced NTG-elevated plasma CGRP, cortical CGRP, and cortical c-Fos.
Adverse eventsnot reported

Exposures

Exposure 1: Cortical TUS at NTG injection / imaging window

Target: cerebral cortex — “cerebral cortex beneath the cranial imaging window (2 mm posterior, 2 mm lateral to bregma)
Device: Olympus / Panametrics · Olympus · V301-SU

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)500✓✓
Pulse duration (ms)50✓✓
Pulse repetition frequency (Hz)1✓✓
Duty cycle (%)5pulse duration × PRF gives 5%✓✓
Sonication duration (s)900✓✓
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
Protocol, in the paper’s words

After NTG (or saline) injection, the NTG+TUS (and Ctrl+TUS) groups received ultrasound stimulation for 15 minutes with a 1 second stimulation interval, targeted at the imaging window; different groups of mice were used for behavioral, optical imaging, and LFP recording sub-experiments.

Flags from extraction

  • n_subjectsGroup sizes (Ctrl+TUS n=6, NTG+TUS n=9) are reused across separate behavioral, optical imaging, and LFP recording sub-experiments using different groups of mice, and a separate n=7 is used for biochemical analysis; the total unique number of TUS-exposed animals is not stated (total across the whole study is 127 mice for all purposes).
  • exposures[0].target.labelThe stimulation site is described only by stereotaxic coordinates relative to bregma ('the image window', -2 mm AP, 2 mm ML), without naming a specific cortical subregion; recorded as generic cerebral cortex.
  • exposures[0].timing.pulse_duration_msPulse duration (50 ms) is derived from the number of cycles per pulse (NC=25000) divided by the fundamental frequency (500 kHz), consistent with duty cycle (5%) / PRF (1 Hz); this conversion is explicitly sanctioned by the extraction instructions.
  • anaesthesiaSurgery for window/electrode implantation was performed under isoflurane anaesthesia, but the paper does not explicitly state whether animals were awake or anaesthetised during the TUS/behavioral/LFP recording sessions themselves; 'awake' is inferred from the freely-moving LFP recording and open-field behavioral testing.