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Acoustic Motor Cortex Stimulation Enhances the Descending Analgesic Pathway to Alleviate Chronic Pain in Mice

Weiliang Fu, Guanghua Yang, Jin Ke, Tianwen Huang, Jinpeng Li, Xiaoyan Chen, Junjie Zou, Zhengrong Lin, Lili Niu, Yongjie Li

IEEE Transactions on Neural Systems and Rehabilitation Engineering 2025 · 10.1109/tnsre.2025.3564033

rodentchronic painbehaviourcellular imaginghistology molecularother mri

Abstract

Chronic pain poses considerable health risks, necessitating the development of effective treatments. Physical modulation of the motor cortex has demonstrated promise for pain relief; however, existing methods require invasive electrode implantation or have limited spatial resolution. Therefore, we developed a non-invasive, high-precision acoustic motor cortex stimulation (aMCS) system to alleviate chronic pain and explore its mechanisms. We developed a wearable aMCS system and employed the spared nerve injury (SNI) method to establish a mouse model of chronic pain. The model mice underwent aMCS with different acoustic parameters, and their pain behaviors were systematically evaluated. Subsequently, we established a long-term spinal cord two-photon system to monitor the effects of aMCS on spinal cord dorsal horn (SCDH) neuronal activity. Next, TRAP2-tdTomato mice were used to examine the effects of aMCS on the motor cortex and other regions of the descending analgesic pathway. Finally, we conducted magnetic resonance imaging, histology, and temperature monitoring to evaluate the safety of aMCS. aMCS with specific parameters significantly ameliorated pain behaviors in a mouse model of chronic pain. Two-photon calcium imaging indicated that aMCS reduced the intensity of neuronal activity in SCDH. Activity mapping in TRAP2-tdTomato mice revealed that aMCS enhanced neuronal activity in the primary motor cortex and zona incerta while diminishing it in the lateral periaqueductal gray and SCDH. Safety assessments confirmed the absence of deleterious effects on the stimulated region. aMCS provides a novel, non-invasive and effective approach to alleviating chronic pain by potentially enhancing the descending analgesic pathway.

Abstract via pubmed.

Speciesmouse (C57BL/6J); mouse (TRAP2-tdTomato)
Subjects8, 7, 6, 7, 5, 3, 4, 3swept animals
Sessions per subjectnot reported
Randomisednot reported
Blindingsingle
Sham / controlinactive transducer
Auditory controlnot reported
Readout timingoffline
Anaesthesiaawake
Readoutsbehaviour, cellular imaging, histology molecular, other mrivon Frey, brush and cold-plate pain behaviour tests; Real-Time Place Avoidance; in vivo two-photon spinal-cord calcium imaging; c-Fos and TRAP2-tdTomato immunohistochemistry; 9.4T MRI, HE/Nissl histology and infrared thermography for safety
Direction of effectbidirectionalaMCS increased neuronal activity (tdTomato labelling) in the primary motor cortex (M1) and zona incerta, but decreased it in the lateral periaqueductal gray and spinal cord dorsal horn (SCDH); SCDH calcium activity was also reduced by two-photon imaging, alongside improved pain behaviours.
Adverse eventsnone observed9.4T MRI showed no significant differences or damage at stimulated sites; HE and Nissl staining showed no tissue damage or neuronal loss; infrared thermography showed no significant temperature alterations at the skull or collimator.

Exposures

Exposure 1: acoustic motor cortex stimulation (aMCS), Groups 1-4 parameter sweep

Target: primary motor cortex — “M1 (primary motor cortex)
Device: custom-built

Pulse timing
Waveformpulsed
Fundamental frequency (kHz)4,170✓✓
Pulse duration (ms)not reported
Pulse repetition frequency (Hz)100, 100, 5, 50swept✓✓
Duty cycle (%)10✓✓
Sonication duration (s)1✓✓
Pressure and intensity, by domain
Free-field pressure (kPa)1,100, 225, 1,100, 1,100swept✓✓
Free-field Isppa (W/cm²)not reported
Free-field Ispta (W/cm²)not reported
In-situ estimatemeasurementsingle value
In-situ pressure (kPa)870, 179, 870, 870swept✓✓
In-situ Isppa (W/cm²)not reported
In-situ Ispta (W/cm²)not reported
Protocol, in the paper’s words

Four parameter groups (paired PRF/pressure combinations, in order Group1-4) shared sonication duration (SD)=1 s, inter-stimulus interval (ISI)=10 s, and duty cycle (DC)=10%. Mice received one week of daily 15-minute aMCS sessions starting on the 8th day post-SNI surgery; sham/control animals had the transducer mounted identically but received no ultrasound energy. Separate cohorts received Group 1 parameters for two-photon spinal calcium imaging (1 week of aMCS), TRAP2 activity mapping (2 consecutive days of 15 min aMCS), and safety assessment.

Flags from extraction

  • n_subjectsPaper never states one overall total of aMCS-exposed mice (95 total mice includes sham/control/excluded animals); listed the per-cohort group sizes that received real aMCS (behaviour Groups 1-4: n=8,7,6,7; SCDH calcium-imaging cohort: n=3; TRAP2 cohort: n=4; safety cohort: n=3) rather than summing them.
  • exposures[0].timing.pulse_duration_msOnly PRF (100/5/50 Hz) and duty cycle (10%) are stated; pulse duration is not independently reported and was not computed from duty_cycle/PRF per the rule against deriving this value.
  • exposures[0].timing.pulse_repetition_frequency_hzPRF and acoustic pressure vary together across four named groups (paired combinations, not a full factorial); recorded as parallel lists in Group1-4 order rather than independent sweeps.
  • n_sessions_per_subjectSession count differs by sub-experiment: ~7 daily sessions (1 week) for the main behaviour and SCDH-imaging cohorts, 2 sessions for the TRAP2 cohort, and an unspecified number for the safety cohort; no single figure applies to the whole study.
  • exposures[0].deviceTransducer described only as custom-built (12 mm radius of curvature, 4.17 MHz); no manufacturer or model name given for the transducer itself (only for the function generator and amplifier).