Parameter-dependent cell-type specific effects of transcranial focused ultrasound stimulation in an awake head-fixed rodent model.

Transcranial focused ultrasound (tFUS) is a promising neuromodulation technique able to target shallow and deep brain structures with high precision. Previous studies have demonstrated that tFUS stimulation responses are cell-type specific, and specifically tFUS can elicit time-locked neural activity in regular spiking units (RSUs) that is sensitive to increases in pulse repetition frequency (PRF), while time-locked responses are not seen in fast spiking units (FSUs). These findings suggest a unique capability of tFUS to alter circuit network dynamics with cell-type specificity; however, these results could be biased by the use of anesthesia, which significantly modulates neural activities.In this study, we developed an awake head-fixed rat model specifically designed for simultaneous tFUS stimulation using a customized 128-element ultrasound array transducer, and recording of spiking data. Using this novel animal model, we examined a series of PRFs and burst duty cycles (DCs) to determine their effects on neuronal subpopulations without anesthesia.We observed cell type specific responses to varying PRF and DC in the awake setting as well as the anesthetized setting, with time locked responses observed in RSU and delayed responses in FSU. Anesthesia broadly was found to dampen responses to tFUS, and affected the latency of delayed responses. Preferred parameters for inducing time-locked responses appear to be 1500 Hz PRF and 60% DC.We conclude that despite some differences in response, isoflurane anesthesia is not a major confound in studying the cell-type specificity of ultrasound neuromodulation, but may affect studies of circuit dynamics and FSU. Our developed awake model will allow for future investigations without this confound.