Viral-Mediated Optogenetic Stimulation of Peripheral Motor Nerves in Non-human Primates.

Objective: Reanimation of muscles paralyzed by disease states such as spinal cord injury remains a highly sought therapeutic goal of neuroprosthetic research. Optogenetic stimulation of peripheral motor nerves expressing light-sensitive opsins is a promising approach to muscle reanimation that may overcome several drawbacks of traditional methods such as functional electrical stimulation (FES). However, the utility of these methods has only been demonstrated in rodents to date, while translation to clinical practice will likely first require demonstration and refinement of these gene therapy techniques in non-human primates. Approach: Three rhesus macaques were injected intramuscularly with either one or both of two optogenetic constructs (AAV6-hSyn-ChR2-eYFP and/or AAV6-hSyn-Chronos-eYFP) to transduce opsin expression in the corresponding nerves. Neuromuscular junctions were targeted for virus delivery using an electrical stimulating injection technique. Functional opsin expression was periodically evaluated up to 13 weeks post-injection by optically stimulating targeted nerves with a 472 nm fiber-coupled laser while recording electromyographic (EMG) responses. Main Results: One monkey demonstrated functional expression of ChR2 at 8 weeks post-injection in each of two injected muscles, while the second monkey briefly exhibited contractions coupled to optical stimulation in a muscle injected with the Chronos construct at 10 weeks. A third monkey injected only in one muscle with the ChR2 construct showed strong optically coupled contractions at 5 ½ weeks which then disappeared by 9 weeks. EMG responses to optical stimulation of ChR2-transduced nerves demonstrated graded recruitment relative to both stimulus pulse-width and light intensity, and followed stimulus trains up to 16 Hz. In addition, the EMG response to prolonged stimulation showed delayed fatigue over several minutes. Significance: These results demonstrate the feasibility of viral transduction of peripheral motor nerves for functional optical stimulation of motor activity in non-human primates, a variable timeline of opsin expression in a animal model closer to humans, and fundamental EMG response characteristics to optical nerve stimulation. Together, they represent an important step in translating these optogenetic techniques as a clinically viable gene therapy.

Multi-scale, multi-modal analysis uncovers complex relationship at the brain tissue-implant neural interface: new emphasis on the biological interface.

OBJECTIVE: Implantable neural electrode devices are important tools for neuroscience research and have an increasing range of clinical applications. However, the intricacies of the biological response after implantation, and their ultimate impact on recording performance, remain challenging to elucidate. Establishing a relationship between the neurobiology and chronic recording performance is confounded by technical challenges related to traditional electrophysiological, material, and histological limitations. This can greatly impact the interpretations of results pertaining to device performance and tissue health surrounding the implant. APPROACH: In this work, electrophysiological activity and immunohistological analysis are compared after controlling for motion artifacts, quiescent neuronal activity, and material failure of devices in order to better understand the relationship between histology and electrophysiological outcomes. MAIN RESULTS: Even after carefully accounting for these factors, the presence of viable neurons and lack of glial scarring does not convey single unit recording performance. SIGNIFICANCE: To better understand the biological factors influencing neural activity, detailed cellular and molecular tissue responses were examined. Decreases in neural activity and blood oxygenation in the tissue surrounding the implant, shift in expression levels of vesicular transporter proteins and ion channels, axon and myelin injury, and interrupted blood flow in nearby capillaries can impact neural activity around implanted neural interfaces. Combined, these tissue changes highlight the need for more comprehensive, basic science research to elucidate the relationship between biology and chronic electrophysiology performance in order to advance neural technologies.

Idle state classification using spiking activity and local field potentials in a brain computer interface.

Previous studies of intracortical brain-computer interfaces (BCIs) have often focused on or compared the use of spiking activity and local field potentials (LFPs) for decoding kinematic movement parameters. Conversely, using these signals to detect the initial intention to use a neuroprosthetic device or not has remained a relatively understudied problem. In this study, we examined the relative performance of spiking activity and LFP signals in detecting discrete state changes in attention regarding a user's desire to actively control a BCI device. Preliminary offline results suggest that the beta and high gamma frequency bands of LFP activity demonstrated a capacity for discriminating idle/active BCI control states equal to or greater than firing rate activity on the same channel. Population classifier models using either signal modality demonstrated an indistinguishably high degree of accuracy in decoding rest periods from active BCI reach periods as well as other portions of active BCI task trials. These results suggest that either signal modality may be used to reliably detect discrete state changes on a fine time scale for the purpose of gating neural prosthetic movements.

Differentiating closed-loop cortical intention from rest: building an asynchronous electrocorticographic BCI.

OBJECTIVE: Recent experiments have shown that electrocorticography (ECoG) can provide robust control signals for a brain-computer interface (BCI). Strategies that attempt to adapt a BCI control algorithm by learning from past trials often assume that the subject is attending to each training trial. Likewise, automatic disabling of movement control would be desirable during resting periods when random brain fluctuations might cause unintended movements of a device. To this end, our goal was to identify ECoG differences that arise between periods of active BCI use and rest. APPROACH: We examined spectral differences in multi-channel, epidural micro-ECoG signals recorded from non-human primates when rest periods were interleaved between blocks of an active BCI control task. MAIN RESULTS: Post-hoc analyses demonstrated that these states can be decoded accurately on both a trial-by-trial and real-time basis, and this discriminability remains robust over a period of weeks. In addition, high gamma frequencies showed greater modulation with desired movement direction, while lower frequency components demonstrated greater amplitude differences between task and rest periods, suggesting possible specialized BCI roles for these frequencies. SIGNIFICANCE: The results presented here provide valuable insight into the neurophysiology of BCI control as well as important considerations toward the design of an asynchronous BCI system.

Cortical adaptation to a chronic micro-electrocorticographic brain computer interface.

Brain-computer interface (BCI) technology decodes neural signals in real time to control external devices. In this study, chronic epidural micro-electrocorticographic recordings were performed over primary motor (M1) and dorsal premotor (PMd) cortex of three macaque monkeys. The differential gamma-band amplitude (75-105 Hz) from two arbitrarily chosen 300 µm electrodes (one located over each cortical area) was used for closed-loop control of a one-dimensional BCI device. Each monkey rapidly learned over a period of days to successfully control the velocity of a computer cursor. While both cortical areas contributed to success on the BCI task, the control signals from M1 were consistently modulated more strongly than those from PMd. Additionally, we observe that gamma-band power during active BCI control is always above resting brain activity. This suggests that purposeful gamma-band modulation is an active process that is obtained through increased cortical activation.

Dynamic control of extracellular environment in in vitro neural recording systems.

A technique is presented for rapid fabrication of microfluidic channels on top of multichannel in vitro neural recording electrode arrays. The channels allow dynamic control of both stable and transient flow patterns over localized areas of the array, over biologically relevant timescales. A cellular model consisting of thermally sensitive dorsal root ganglion neurons was integrated into the devices. The device was used to demonstrate precise control of the extracellular microenvironment of individual cells on the array. Since the methods presented here are not specific to a particular cell type or neural recording system, the technique is amenable to a wide range of applications within the neuroscience field.