Real-time Monitoring Unveils Three Distinct Neuronal Response Patterns to SAW Ultrasound via L-type Calcium Channels
Yiming Chen, Wenxu Tang, Yifan Wang, Ya Gao, Jiaqi Hu, Yixuan Lu, Long Meng, Hairong Zheng, Yi Feng, Liming Cheng, Wenyong Fan, Qian Cheng, Lei Xue
Neuroscience Bulletin 2026 · 10.1007/s12264-025-01457-6
Abstract
Ultrasound neuromodulation shows promise for treating neurological disorders, but the underlying mechanisms remain unclear. Here, we developed an integrated surface acoustic wave (SAW) ultrasound chip enabling simultaneous electrophysiological recording and Ca 2+ imaging of cultured hippocampal neurons to investigate neuronal excitability and synaptic transmission during ultrasound stimulation. This study revealed, for the first time, three distinct neuronal response patterns induced by SAW ultrasound: an immediate response showing rapid activation, a delayed response exhibiting facilitation after several minutes, and a non-response maintaining baseline activity. Ultrasound stimulation increased action potential firing, enhanced excitatory postsynaptic currents, and elevated intracellular Ca 2+ levels. These effects were dependent on extracellular Ca 2+ influx and primarily dominated by L-type Ca 2+ channels. Our findings suggest that individual neurons exhibit heterogeneous responses to SAW ultrasound stimulation based on their intracellular Ca 2+ levels and L-type Ca 2+ channel activity. This integrated approach provides new insights into the cellular mechanisms of ultrasound neuromodulation while highlighting the potential of SAW technology for precise, cell-type-specific neural control.
Abstract via europepmc.
Exposures
Exposure 1: SAW ultrasound chip, cultured rat hippocampal neurons, 7.65, 11.87 and 15.88 MHz
Target: cultured neurons — “cultured hippocampal neurons”
Device: custom-built · SAW US neuromodulation chip, interdigital transducers on LiTaO3 ✓
| Waveform | continuous | |
|---|---|---|
| Fundamental frequency (kHz) | 7,650, 11,870, 15,880swept | ✓✓✓⚑ |
| Pulse duration (ms) | not applicable | |
| Pulse repetition frequency (Hz) | not applicable | |
| Duty cycle (%) | not applicable | |
| Sonication duration (s) | 60, 120swept | ✓✓✓⚑ |
| Free-field pressure (kPa) | not reported | |
|---|---|---|
| Free-field Isppa (W/cm²) | not reported | |
| Free-field Ispta (W/cm²) | not reported | |
| In-situ estimate | not applicable | |
| In-situ pressure (kPa) | not applicable | |
| In-situ Isppa (W/cm²) | not applicable | |
| In-situ Ispta (W/cm²) | not applicable | |
| Ispta, domain unspecified (W/cm²) | 0.025, 0.4swept | ✓✓✓⚑ |
Continuous standing SAW US; 60-s sessions at 120-s intervals at increasing Ispta (0, 25, 50, 100, 400 mW/cm2) for each frequency; later experiments used 2-min stimulation at 100 mW/cm2 within a 20-min recording.
Flags from extraction
exposures[0].unspecified_domain.ispta_w_cm2— Ispta measured by laser Doppler velocimetry on the chip, in mW/cm2 (0 to 400); converted to W/cm2; domain not a water or in-situ statement.n_subjects— 38 is the number of neurons recorded in the temporal-dynamics experiment; other experiments use different n. No total culture count given.exposures[0].timing.sonication_duration_s— 60 s used in the frequency/intensity sweep; later experiments used 2-min stimulation.exposures[0].fundamental_frequency_khz— Measured resonant frequencies 7.65, 11.87, 15.88 MHz (design 8, 12, 16 MHz).exposures[0].device— Chips designed and fabricated by the authors; surface acoustic wave rather than bulk transducer.