Focused ultrasound stimulation of an ex-vivo Aplysia abdominal ganglion preparation.

Background: A growing body of research demonstrates that focused ultrasound stimulates activity in human and other mammalian nervous systems. However, there is no consensus on which sonication parameters are optimal. Furthermore, the mechanism of action behind ultrasound neurostimulation remains poorly understood. An invertebrate model greatly reduces biological complexity, permitting a systematic evaluation of sonication parameters suitable for ultrasound neurostimulation.

New Method: Here, we describe the use of focused ultrasound stimulation with an ex-vivo abdominal ganglion preparation of the California sea hare, Aplysia californica, a long-standing model system in neurobiology. We developed a system for stimulating an isolated ganglion preparation while obtaining extracellular recordings from nerves. The focused ultrasound stimulation uses one of two single-element transducers, enabling stimulation at four distinct carrier frequencies (0.515 MHz, 1.l MHz, 1.61 MHz, 3.41 MHz).

Results: Using continuous wave ultrasound, we stimulated the ganglion at all four frequencies, and we present quantitative evaluation of elicited activation at four different sonication durations and three peak pressure levels, eliciting up to a 57-fold increase in spiking frequency.

Comparison With Electrical Stimulation: We demonstrated that ultrasound-induced activation is repeatable, and the response consistency is comparable to electrical stimulation.

Conclusions: Due to the relative ease of long-term recordings for many hours, this ex-vivo ganglion preparation is suitable for investigating sonication parameters and the effects of focused ultrasound stimulation on neurons.