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Transcranial ultrasound neuromodulation facilitates isoflurane-induced general anesthesia recovery and improves cognition in mice

Jiaru He, Yiyue Zhu, Canwen Wu, Junwei Wu, Yan Chen, Maodan Yuan, Zhongwen Cheng, Lvming Zeng, Xuanrong Ji

Ultrasonics 2023, 135, 107132 · 10.1016/j.ultras.2023.107132

rodenthealthyhistology molecularbehaviourother

Abstract

Delayed arousal and cognitive dysfunction are common, especially in older patients after general anesthesia (GA). Elevating central nervous system serotonin (5-HT) levels can promote recovery from GA and increase synaptic plasticity to improve cognition. Ultrasound neuromodulation has become a noninvasive physical intervention therapy with high spatial resolution and penetration depth, which can modulate neuronal excitability to treat psychiatric and neurodegenerative diseases. This study aims to use ultrasound to noninvasively modulate the brain 5-HT levels of mice to promote recovery from GA and improve cognition in mice. The dorsal raphe nucleus (DRN) of mice during GA was stimulated by the 1.1 MHz ultrasound with a negative pressure of 356 kPa, and the liquid chromatography coupled tandem mass spectrometry (LC-MS/MS) method was used to measure the DRN 5-HT concentrations. The mice's recovery time from GA was assessed, and the cognition was evaluated through spontaneous alternation Y-maze and novel object recognition (NOR) tests. After ultrasound stimulation, the mice's DRN 5-HT levels were significantly increased (control: 554.0 ± 103.2 ng/g, anesthesia + US: 664.2 ± 84.1 ng/g, *p = 0.0389); the GA recovery time (return of the righting reflex (RORR) emergence latency time) of mice was significantly reduced (anesthesia: 331.6 ± 70 s, anesthesia + US: 223.2 ± 67.7 s, *p = 0.0215); the spontaneous rotation behavior score of mice was significantly increased (anesthesia: 59.46 ± 5.26 %, anesthesia + US: 68.55 ± 5.24 %; *p = 0.0126); the recognition index was significantly increased (anesthesia: 55.02 ± 6.23 %, anesthesia + US: 78.52 ± 12.21 %; ***p = 0.0009). This study indicates that ultrasound stimulation of DRN increases serotonin levels, accelerates recovery from anesthesia, and improves cognition, which could be an important strategy for treating delayed arousal, postoperative delirium, or even lasting cognitive dysfunction after GA.

Abstract via europepmc.

Speciesmouse
Subjects4, 4, 4, 4, 10, 8, 8swept animals
Sessions per subject1
Randomisedyes
Blindingdouble
Sham / controlundescribed
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutshistology molecular, behaviour, otherc-Fos immunohistochemistry; H&E histology; LC-MS/MS quantification of DRN serotonin (5-HT); return-of-righting-reflex (RORR) and other emergence-latency measures from general anaesthesia; Y-maze spontaneous alternation; novel object recognition (NOR) test
Direction of effectexcitatoryUltrasound stimulation of the DRN increased c-Fos expression and 5-HT tissue levels, accelerated emergence from isoflurane anaesthesia (reduced RORR latency), and improved spontaneous-alternation and novel-object-recognition performance; effects on GA recovery and cognition were reduced by the 5-HT2A antagonist M100907.
Adverse eventsnone observedH&E staining of the DRN region showed no obvious microhemorrhage, edema, or cell necrosis in ultrasound-stimulated versus sham mice.

Exposures

Exposure 1: Dorsal raphe nucleus (DRN) stimulation, 1.1 MHz

Target: raphe nuclei — “dorsal raphe nucleus (DRN)
Device: custom-built

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,100✓✓
Pulse duration (ms)0.5✓✓
Pulse repetition frequency (Hz)1,000✓✓
Duty cycle (%)50pulse duration × PRF gives 50%✓✓
Sonication duration (s)1✓✓
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 estimatemeasurementsingle value
In-situ pressure (kPa)356, 500, 714swept✓✓
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Low-intensity pulsed ultrasound delivered to the DRN with a 1-second interstimulus interval between 1-second sonication trains, applied during the last 40 minutes of a 1-hour 2.0% isoflurane anaesthesia session (c-Fos and 5-HT-level experiments used the same 40-minute stimulation window; the 5-HT2A antagonist experiment also used 356 kPa for 40 minutes).

Flags from extraction

  • n_subjectsStudy comprises several separate sub-experiments (c-Fos, dose-response 5-HT, GA-recovery/cognition, 5-HT2A antagonist) each with its own ultrasound-exposed group(s); no single total across the whole study is stated, so exposed-group sizes are listed rather than summed (86 mice were used in total across the whole study, but not all received ultrasound).
  • sham_typeThe sham/control groups in each sub-experiment received no ultrasound at all during the anaesthesia window; the paper does not state whether an inactive transducer was placed on the head for these animals, so the sham mechanism is not fully described.
  • n_sessions_per_subjectEach animal appears to receive a single stimulation session per sub-experiment, but this is not explicitly stated as a session count.