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The cerebellum takes in a great deal of sensory information from the periphery and descending signals from the cerebral cortices. It has been debated whether the paramedian lobule (PML) in the rat and its paravermal regions that project to the interpositus nucleus (IPN) are primarily involved in motor execution or motor planning. Studies that have relied on single spike recordings in behaving animals have led to conflicting conclusions regarding this issue. In this study, we tried a different approach and investigated the correlation of field potentials and multi-unit signals recorded with multi-electrode arrays from the PML cortex along with the forelimb electromyography (EMG) signals in rats during behavior. Linear regression was performed to predict the EMG signal envelopes using the PML activity for various time shifts (±25, ±50, ±100, and ± 400 ms) between the two signals to determine a causal relation. The highest correlations (~0.5 on average) between the neural and EMG envelopes were observed for zero and small (±25 ms) time shifts and decreased with larger time shifts in both directions, suggesting that paravermal PML is involved both in processing of sensory signals and motor execution in the context of forelimb reaching behavior. EMG envelopes were predicted with higher success rates when neural signals from multiple phases of the behavior were utilized for regression. The forelimb extension phase was the most difficult to predict while the releasing of the bar phase prediction was the most successful. The high frequency (>300 Hz) components of the neural signal, reflecting multi-unit activity, had a higher contribution to the EMG prediction than did the lower frequency components, corresponding to local field potentials. The results of this study suggest that the paravermal PML in the rat cerebellum is primarily involved in the execution of forelimb movements rather than the planning aspect and that the PML is more active at the initiation and termination of the behavior, rather than the progression.

Transcranial alternating current stimulation (tACS) is a non-invasive neuromodulation technique that is being tested clinically for treatment of a variety of neural disorders. Animal studies investigating the underlying mechanisms of tACS are scarce, and nearly absent in the cerebellum. In the present study, we applied 10–400 Hz alternating currents (AC) to the cerebellar cortex in ketamine/xylazine anesthetized rats. The spiking activity of cerebellar nuclear (CN) cells was transsynaptically entrained to the frequency of AC stimulation in an intensity and frequency-dependent manner. Interestingly, there was a tuning curve for modulation where the frequencies in the midrange (100 and 150 Hz) were more effective, although the stimulation frequency for maximum modulation differed for each CN cell with slight dependence on the stimulation amplitude. CN spikes were entrained with latencies of a few milliseconds with respect to the AC stimulation cycle. These short latencies and that the transsynaptic modulation of the CN cells can occur at such high frequencies strongly suggests that PC simple spike synchrony at millisecond time scales is the underlying mechanism for CN cell entrainment. These results show that subthreshold AC stimulation can induce such PC spike synchrony without resorting to supra-threshold pulse stimulation for precise timing. Transsynaptic entrainment of deep CN cells via cortical stimulation could help keep stimulation currents within safety limits in tACS applications, allowing development of tACS as an alternative treatment to deep cerebellar stimulation. Our results also provide a possible explanation for human trials of cerebellar stimulation where the functional impacts of tACS were frequency dependent.

IntroductionCerebellar transcranial alternating current stimulation (ctACS) has shown promise as a therapeutic modality for treating a variety of neurological disorders, and for affecting normal learning processes. Yet, little is known about how electric fields induced by applied currents affect cerebellar activity in the mammalian cerebellum under in vivo conditions.MethodsAlternating current (AC) stimulation with frequencies from 0.5 to 20 Hz was applied to the surface of the cerebellum in anesthetized rats. Extracellular recordings were obtained from Purkinje cells (PC), cerebellar and vestibular nuclear neurons, and other cerebellar cortical neurons.Results and discussionAC stimulation modulated the activity of all classes of neurons. Cerebellar and vestibular nuclear neurons most often showed increased spike activity during the negative phase of the AC stimulation. Purkinje cell simple spike activity was also increased during the negative phase at most locations, except for the cortex directly below the stimulus electrode, where activity was most often increased during the positive phase of the AC cycle. Other cortical neurons showed a more mixed, generally weaker pattern of modulation. The patterns of Purkinje cell responses suggest that AC stimulation induces a complex electrical field with changes in amplitude and orientation between local regions that may reflect the folding of the cerebellar cortex. Direct measurements of the induced electric field show that it deviates significantly from the theoretically predicted radial field for an isotropic, homogeneous medium, in both its orientation and magnitude. These results have relevance for models of the electric field induced in the cerebellum by AC stimulation.

Abstract Background Electrocorticography (ECoG) arrays are commonly used to record the brain activity both in animal and human subjects. There is a lack of guidelines in the literature as to how the array geometry, particularly the via holes in the substrate, affects the recorded signals. A finite element (FE) model was developed to simulate the electric field generated by neurons located at different depths in the rat brain cortex and a micro ECoG array (μECoG) was placed on the pia surface for recording the neural signal. The array design chosen was a typical array of 8 × 8 circular (100 μm in diam.) contacts with 500 μm pitch. The size of the via holes between the recording contacts was varied to see the effect. Results The results showed that recorded signal amplitudes were reduced if the substrate was smaller than about four times the depth of the neuron in the gray matter. The signal amplitude profiles had dips around the via holes and the amplitudes were also lower at the contact sites as compared to the design without the holes; an effect that increased with the hole size. Another noteworthy result is that the spatial selectivity of the multi-contact recordings could be improved or reduced by the selection of the via hole sizes, and the effect depended on the distance between the neuron pair targeted for selective recording and its depth. Conclusions The results suggest that the via-hole size clearly affects the recorded neural signal amplitudes and it can be leveraged as a parameter to reduce the inter-channel correlation and thus maximize the information content of neural signals with μECoG arrays.

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.

Transcranial electrical stimulation (tES) is rapidly becoming an indispensable clinical tool with its different forms. Animal data are crucially needed for better understanding of the underlying mechanisms of tES. For reproducibility of results in animal experiments, the electric fields (E-Fields) inside the brain parenchyma induced by the injected currents need to be predicted accurately. In this study, we measured the electrical fields in the rat brain perpendicular to the brain surface, i.e. vertical electric field (VE-field), when the stimulation electrode was placed over the skin, skull, or dura mater through a craniotomy hole. The E-field attenuation through the skin was a few times larger than that of the skull and the presence of skin substantially reduced the VE-field peak at the cortical surface near the electrode. The VE-field declined much quicker in the gray matter underneath the pial surface than it did in the white matter, and thus the large VE-fields were contained mostly in the gray matter. The transition at the gray/white matter border caused a significant peak in the VE-field, as well as at other local inhomogeneties. A conductivity value of 0.57 S/m is predicted as a global value for the whole brain by matching our VE-field measurements to the field profile given by analytical equations for volume conductors. Finally, insertion of the current return electrode into the shoulder, submandibular, and hind leg muscles had virtually no effects on the measured E-field amplitudes in the cortex underneath the epidural electrodes.

Advanced algorithms are required to reveal the complex relations between neural and behavioral data. In this study, forelimb electromyography (EMG) signals were reconstructed from multi-unit neural signals recorded with multiple electrode arrays (MEAs) from the corticospinal tract (CST) in rats. A six-layer convolutional neural network (CNN) was compared with linear decoders for predicting the EMG signal. The network contained three session-dependent Rectified Linear Unit (ReLU) feature layers and three Gamma function layers were shared between sessions. Coefficient of determination (R2) values over 0.2 and correlations over 0.5 were achieved for reconstruction within individual sessions in multiple animals, even though the forelimb position was unconstrained for most of the behavior duration. The CNN performed visibily better than the linear decoders and model responses outlasted the activation duration of the rat neuromuscular system. These findings suggest that the CNN model implicitly predicted short-term dynamics of skilled forelimb movements from neural signals. These results are encouraging that similar problems in neural signal processing may be solved using variants of CNNs defined with simple analytical functions. Low powered firmware can be developed to house these CNN solutions in real-time applications.

Extracellular neural recording, with multi-electrode arrays (MEAs), is a powerful method used to study neural function at the network level. However, in a high density array, it can be costly and time consuming to integrate the active circuit with the expensive electrodes. In this paper, we present a 4 mm × 4 mm neural recording integrated circuit (IC) chip, utilizing IBM C4 bumps as recording electrodes, which enable a seamless active chip and electrode integration. The IC chip was designed and fabricated in a 0.13 μm BiCMOS process for both in vitro and in vivo applications. It has an input-referred noise of 4.6 μV rms for the bandwidth of 10 Hz to 10 kHz and a power dissipation of 11.25 mW at 2.5 V, or 43.9 μW per input channel. This prototype is scalable for implementing larger number and higher density electrode arrays. To validate the functionality of the chip, electrical testing results and acute in vivo recordings from a rat barrel cortex are presented.

A multi-electrode array (MEA) was implanted in the dorsolateral funiculus of the cervical spinal cord to record descending information during behavior in freely moving rats. Neural signals were characterized in terms of frequency and information content. Frequency analysis revealed components both at the range of local field potentials and multi-unit activity. Coherence between channels decreased steadily with inter-contact distance and frequency suggesting greater spatial selectivity for multi-unit activity compared to local field potentials. Principal component analysis (PCA) extracted multiple channels of neural activity with patterns that correlated to the behavior, indicating multiple dimensionality of the signals. Two different behaviors involving the forelimbs, face cleaning and food reaching, generated neural signals through distinctly different combination of neural channels, which suggested that these two behaviors could readily be differentiated from recordings. This preliminary data demonstrated that descending spinal cord signals recorded with MEAs can be used to extract multiple channels of command control information and potentially be utilized as a means of communication in high level spinal cord injury subjects.