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Modulation of rapid eye movement sleep deprivation via transcranial ultrasound stimulation of the basal forebrain in mice

Yue Wu, Haiqing Wang, Mengke Wang, Pengyu Ren, Siyuan Zhang

Ultrasonics 2026, 108282 · 10.1016/j.ultras.2026.108282

rodentothereeg megbehaviourhistology molecularother

Abstract

This study presents a strategy of non‑invasive transcranial ultrasound stimulation (TUS) to mitigate sleep insufficiency. Mice were subjected to 20 h of sleep deprivation, while receiving 10 min of TUS of the basal forebrain for 7 days. Sleep architecture was evaluated by electroencephalographic analysis, while anxiety-like behavior and cognitive performance were assessed using open field testing and Y-maze. Hippocampal neuronal integrity was examined by Nissl staining, and serum cortisol levels was assessed by ELISA, whereas basal forebrain cholinergic activation was analyzed using c-Fos immunofluorescence. TUS significantly increased rapid eye movement (REM) sleep compared with the sleep-deprived group (14.97 ± 1.05% vs. 10.86 ± 1.11%, P < 0.05), while non-rapid eye movement-related delta power rose to 130.77% of normal control levels (P < 0.0001) and REM-related theta power was enhanced (P < 0.05). TUS improved cognitive performance, increasing Y-maze alternation rate to 61.42 ± 1.31% versus 50.05 ± 1.59% in sleep-deprived mice (P < 0.001). Histological analysis demonstrated marked neuroprotection, with TUS significantly restoring hippocampal neuronal counts in the cornu ammonis 1 and dentate gyrus. In addition, serum cortisol levels were elevated toward normal values (P < 0.05), accompanied by increased c-Fos expression, suggesting enhanced basal forebrain neuronal activity and potential cholinergic involvement. These findings suggest that TUS provides a novel non-pharmacological treatment strategy targeting the basal forebrain in the sleep deprivation mouse model.

Abstract via europepmc.

Speciesmouse (C57BL/6N)
Subjects8 animals
Sessions per subject7
Randomisedyes
Blindingnot reported
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingoffline
Anaesthesiaanaesthetised
Readoutseeg meg, behaviour, histology molecular, otherY-maze spontaneous alternation; open field test; Nissl staining of hippocampal neuron counts; c-Fos immunofluorescence; serum cortisol ELISA
Direction of effectexcitatoryTUS of the basal forebrain increased c-Fos expression (neuronal activation marker) in the BF, increased REM sleep percentage and NREM delta / REM theta power toward or above normal-control levels, and improved cognitive/behavioural and cortisol measures relative to sleep-deprived mice.
Adverse eventsnot reported

Exposures

Exposure 1: TUS of the bilateral medial basal forebrain in sleep-deprived mice

Target: basal forebrain — “bilateral medial basal forebrain (AP = +0.24 mm, ML = ±0.25 mm, DV = -5.00 mm relative to bregma)
Device: not reported

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)1,000✓✓
Pulse duration (ms)2✓✓
Pulse repetition frequency (Hz)250✓✓
Duty cycle (%)50pulse duration × PRF gives 50%✓✓
Sonication duration (s)0.2✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)270✓✓
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

The total sonication duration (TSD) was 200 ms, with an inter-stimulation interval (ISI) of 5 s. The US group received ultrasound stimulation for 10 min per day while the mice were sleep deprived, for 7 consecutive days.

Flags from extraction

  • n_subjectsMain group-design statement gives n=8 per group, but several figure legends report smaller subsample sizes for specific assays (e.g., n=5 for EEG/behaviour, n=3 for cortisol); n=8 (the ultrasound-stimulated US group) is used here.
  • exposures[0].deviceThe transducer is described only as 'a small 1 MHz focused transducer' with no manufacturer or model given.
  • exposures[0].free_field.pressure_kpaOnly negative peak pressure and mechanical index are reported; no ISPPA/ISPTA intensity values are given.