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Background Focused ultrasound (FUS) has excellent characteristics over other non-invasive stimulation methods in terms of spatial resolution and steering capability of the target. FUS has not been tested in the cerebellar cortex and cellular effects of FUS are not fully understood. Objective/hypothesis To investigate how the activity of cerebellar Purkinje cells (PCs) is modulated by FUS with varying pulse durations and pulse repetition frequencies. Methods A glass microelectrode was inserted into the cerebellar vermis lobule 6 from the dorsal side to extracellularly record single unit activity of the PCs in anesthetized rats. Ultrasonic stimulation (500 kHz) was applied through a coupling cone, filled with degassed water, from the posterior side to target the recording area with varying pulse durations and frequencies. Results Simple spike (SS) activity of PCs was entrained by the FUS pattern where the probability of spike occurrences peaked at around 1 ms following the onset of the stimulus regardless of its duration (0.5, 1, or 2 ms). The level of entrainment was stronger with shorter pulse durations at 50-Hz pulse repetition frequency (PRF), however, peri-event histograms spread wider and the peaks delayed slightly at 100-Hz PRF, suggesting involvement of a long-lasting inhibitory mechanism. There was no significant difference between the average firing rates in the baseline and stimulation periods. Conclusion FUS can entrain spiking activity of single cells on a spike-by-spike basis as demonstrated here in the rat cerebellar cortex. The observed modulation potentially results from the aggregate of excitatory and inhibitory effects of FUS on the entire cortical network rather than on the PCs alone.

Background Transcranial electrical stimulation (tES) shows promise to treat neurological disorders. Knowledge of how the orthogonal components of the electric field (E-field) alter neuronal activity is required for strategic placement of transcranial electrodes. Yet, essentially no information exists on this relationship for mammalian cerebellum in vivo, despite the cerebellum being a target for clinical tES studies. Objective To characterize how cerebellar Purkinje cell (PC) activity varies with the intensity, frequency, and direction of applied AC and DC E-fields. Methods Extracellular recordings were obtained from vermis lobule 7 PCs in anesthetized rats. AC (2–100 Hz) or DC E-fields were generated in a range of intensities (0.75–30 mV/mm) in three orthogonal directions. Field-evoked PC simple spike activity was characterized in terms of firing rate modulation and phase-locking as a function of these parameters. t-tests were used for statistical comparisons. Results The effect of applied E-fields was direction and intensity dependent, with rostrocaudally directed fields causing stronger modulations than dorsoventral fields and mediolaterally directed ones causing little to no effect, on average. The directionality dependent modulation suggests that PC is the primary cell type affected the most by electric stimulation, and this effect was probably given rise by a large dendritic tree and a soma. AC stimulation entrained activity in a frequency dependent manner, with stronger phase-locking to the stimulus cycle at higher frequencies. DC fields produced a modulation consisting of strong transients at current onset and offset with an intervening plateau. Conclusion Orientation of the exogenous E-field critically determines the modulation depth of cerebellar cortical output. With properly oriented fields, PC simple spike activity can strongly be entrained by AC fields, overriding the spontaneous firing pattern.

Associative plasticity occurs when two stimuli converge on a common neural target. Previous efforts to promote associative plasticity have targeted cortex, with variable and moderate effects. In addition, the targeted circuits are inferred, rather than tested directly. In contrast, we sought to target the strong convergence between motor and sensory systems in the spinal cord. We developed spinal cord associative plasticity, precisely timed pairing of motor cortex and dorsal spinal cord stimulations, to target this interaction. We tested the hypothesis that properly timed paired stimulation would strengthen the sensorimotor connections in the spinal cord and improve recovery after spinal cord injury. We tested physiological effects of paired stimulation, the pathways that mediate it, and its function in a preclinical trial. Subthreshold spinal cord stimulation strongly augmented motor cortex evoked muscle potentials at the time they were paired, but only when they arrived synchronously in the spinal cord. This paired stimulation effect depended on both cortical descending motor and spinal cord proprioceptive afferents; selective inactivation of either of these pathways fully abrogated the paired stimulation effect. Spinal cord associative plasticity, repetitive pairing of these pathways for 5 or 30 min in awake rats, increased spinal excitability for hours after pairing ended. To apply spinal cord associative plasticity as therapy, we optimized the parameters to promote strong and long-lasting effects. This effect was just as strong in rats with cervical spinal cord injury as in uninjured rats, demonstrating that spared connections after moderate spinal cord injury were sufficient to support plasticity. In a blinded trial, rats received a moderate C4 contusive spinal cord injury. Ten days after injury, they were randomized to 30 min of spinal cord associative plasticity each day for 10 days or sham stimulation. Rats with spinal cord associative plasticity had significantly improved function on the primary outcome measure, a test of dexterity during manipulation of food, at 50 days after spinal cord injury. In addition, rats with spinal cord associative plasticity had persistently stronger responses to cortical and spinal stimulation than sham stimulation rats, indicating a spinal locus of plasticity. After spinal cord associative plasticity, rats had near normalization of H-reflex modulation. The groups had no difference in the rat grimace scale, a measure of pain. We conclude that spinal cord associative plasticity strengthens sensorimotor connections within the spinal cord, resulting in partial recovery of reflex modulation and forelimb function after moderate spinal cord injury. Since both motor cortex and spinal cord stimulation are performed routinely in humans, this approach can be trialled in people with spinal cord injury or other disorders that damage sensorimotor connections and impair dexterity.

Transcranial electrical stimulation (tES) techniques have garnered significant interest due to their non-invasiveness and potential to offer a treatment option in a wide variety of brain disorders. Among several modulation techniques, transcranial alternating current stimulation (tACS) is favored for its ability to entrain the neural oscillations. The cerebellum is one of the targeted sites because of its involvement in motor and cognitive functions. However, animal studies are lacking in the literature looking into the mechanism of action in cerebellar tACS. In this study, we used a rat model and monitored the activity of the cerebellar cortex, which sculpts the cerebellar output by adjusting the firing rate and timing of the neurons in the deep cerebellar nuclei (DCN). For neural recording, a tungsten electrode was inserted into the cerebellar cortex through a craniotomy hole located over the right paramedian lobule (PML). A helical Ag/AgCl wire electrode was placed atop the skull near the caudal edge to inject a 1 Hz biphasic sinusoidal current. Our results showed that the multiunit activity (MUA) of the cerebellar cortex was strongly modulated by tACS. The negative phase of the electric current enhanced the neural firing rate while the positive phase suppressed the activity. Furthermore, the spike rate showed modulation by the instantaneous strength of the injected current within the sinusoidal cycle. This warrants research to further look into the mechanism of tACS acting on the cerebellar cortex at the cellular level.

Understanding the mechanisms underlying traumatic neural injury and the sequelae of events in the acute phase is important for deciding on the best window of therapeutic intervention. We hypothesized that evoked potentials (EP) recorded from the cerebellar cortex can detect mild levels of neural trauma and provide a qualitative assessment tool for progression of cerebellar injury in time. The cerebellar local field potentials evoked by a mechanical tap on the hand and collected with chronically implanted micro-ECoG arrays on the rat cerebellar cortex demonstrated substantial changes both in amplitude and timing as a result of blast-wave induced injury. The results revealed that the largest EP changes occurred within the first day of injury, and partial recoveries were observed from day-1 to day-3, followed by a period of gradual improvements (day-7 to day-14). The mossy fiber (MF) and climbing fiber (CF) mediated components of the EPs were affected differentially. The behavioral tests (ladder rung walking) and immunohistological analysis (calbindin and caspase-3) did not reveal any detectable changes at these blast pressures that are typically considered as mild (100–130 kPa). The results demonstrate the sensitivity of the electrophysiological method and its use as a tool to monitor the progression of cerebellar injuries in longitudinal animal studies.