Synchronous Measurements of Extracellular Action Potentials and Neurochemical Activity with Carbon Fiber Electrodes in Nonhuman Primates.

Measuring the dynamic relationship between neuromodulators, such as dopamine, and neuronal action potentials is imperative to understand how these fundamental modes of neural signaling interact to mediate behavior. We developed methods to measure concurrently dopamine and extracellular action potentials (i.e., spikes) in monkeys. Standard fast-scan cyclic voltammetric (FSCV) electrochemical (EChem) and electrophysiological (EPhys) recording systems are combined and used to collect spike and dopamine signals, respectively, from an array of carbon fiber (CF) sensors implanted in the monkey striatum. FSCV requires the application of small voltages at the implanted sensors to measure redox currents generated from target molecules, such as dopamine. These applied voltages create artifacts at neighboring EPhys-measurement sensors which may lead to misclassification of these signals as physiological spikes. Therefore, simple automated temporal interpolation algorithms were designed to remove these artifacts and enable accurate spike extraction. We validated these methods using simulated artifacts and demonstrated an average spike recovery rate of 84.5%. We identified and discriminated cell-type specific units in the monkey striatum that were shown to correlate to specific behavioral task parameters related to reward size and eye-movement direction. Synchronously recorded spike and dopamine signals displayed contrasting relations to the task variables, suggesting a complex relationship between these two modes of neural signaling. Future application of our methods will help advance our understanding of the interactions between neuromodulator signaling and neuronal activity, to elucidate more detailed mechanisms of neural circuitry and plasticity mediating behaviors in health and in disease.Significance statement We present a simple method for recording synchronous molecular and neuronal spike signals. Conventional electrophysiological and electrochemical instruments are combined without the need for additional hardware. A custom-designed algorithm was made and validated for extracting neuronal action potential signals with high fidelity. We were able to compute cell-type specific spike activity along with molecular dopamine signals related to reward and movement behaviors from measurements made in the monkey striatum. Such combined measurements of neurochemical and extracellular action potentials may help pave the way to elucidating mechanisms of plasticity, and how neuromodulators and neurons are orchestrated to mediate behavior.

An open-source MRI compatible frame for multimodal presurgical mapping in macaque and capuchin monkeys.

BACKGROUND: High-precision neurosurgical targeting in nonhuman primates (NHPs) often requires presurgical anatomical mapping with noninvasive neuroimaging techniques (MRI, CT, PET), allowing for translation of individual anatomical coordinates to surgical stereotaxic apparatus. Given the varied tissue contrasts that these imaging techniques produce, precise alignment of imaging-based coordinates to surgical apparatus can be cumbersome. MRI-compatible stereotaxis with radiopaque fiducial markers offer a straight-forward and reliable solution, but existing commercial options do not fit in conformal head coils that maximize imaging quality. NEW METHOD: We developed a compact MRI-compatible stereotaxis suitable for a variety of NHP species (Macaca mulatta, Macaca fascicularis, and Cebus apella) that allows multimodal alignment through technique-specific fiducial markers. COMPARISON WITH EXISTING METHODS: With the express purpose of compatibility with clinically available MRI, CT, and PET systems, the frame is no larger than a human head, while allowing for imaging NHPs in the supinated position. This design requires no marker implantation, special software, or additional knowledge other than the operation of a common large animal stereotaxis. RESULTS: We demonstrated the applicability of this 3D-printable apparatus across a diverse set of experiments requiring presurgical planning: 1) We demonstrate the accuracy of the fiducial system through a within-MRI cannula insertion and subcortical injection of a viral vector. 2) We also demonstrated accuracy of multimodal (MRI and CT) alignment and coordinate transfer to guide a surgical robot electrode implantation for deep-brain electrophysiology. CONCLUSIONS: The computer-aided design files and engineering drawings are publicly available, with the modular design allowing for low cost and manageable manufacturing.

The prevalence of chronic pain in adolescents in Central Switzerland: A cross- sectional school-based study protocol.

BACKGROUND: Chronic pain is associated with substantial personal suffering and societal costs and is a growing healthcare concern worldwide. While chronic pain has been extensively studied in adults, limited data exists on its prevalence and impact in adolescents. Understanding the prevalence and impact of chronic pain and pain beliefs in adolescents is crucial for developing effective prevention and treatment strategies. This study aims to estimate the prevalence, characteristics, and impact of chronic pain, and explore adolescents' knowledge and beliefs about pain. METHODS: This is an observational cohort study of school-going adolescents aged 11 to 17 years in Central Switzerland. The study will estimate the point prevalence, characteristics (location, intensity, frequency, duration) and impact (PROMIS Pediatric Short Form v2.0 -Pain Interference Scale, PPIS) of chronic pain in school-going adolescents. We will also measure and investigate pupils' beliefs about pain (Concept of Pain Inventory (COPI)). Data will be collected through manual and digital self-report questionnaires and from participants in primary, secondary, and high schools between September 2023 and January 2024. ANALYSES: The primary analyses will utilise descriptive statistics to estimate the point prevalence, characteristics, and impact of chronic pain. Secondary analyses will analyse associations and correlations between chronic pain, impact of pain and beliefs about pain. OUTCOMES: This study will provide an estimate of the prevalence, characteristics and impact of chronic pain in adolescents in Central Switzerland and a measure of adolescents' understanding and beliefs about pain. In doing so, this study will provide insights into the scale of chronic pain as a public health concern. By understanding adolescents' pain beliefs and their influence on pain experience, this study can contribute to the development of educational approaches to enhance adolescents' knowledge and understanding of pain in order to optimise the prevention and treatment of chronic pain in adolescents. The findings may be useful to healthcare professionals and funders, policymakers, and researchers involved in the prevention, assessment, and treatment of pain in adolescents.

Scalable, flexible carbon fiber electrode thread arrays for three-dimensional probing of neurochemical activity in deep brain structures of rodents.

We developed a flexible "electrode-thread" array for recording dopamine neurochemicals from a lateral distribution of subcortical targets (up to 16) transverse to the axis of insertion. Ultrathin (∼10 µm diameter) carbon fiber (CF) electrode-threads (CFETs) are clustered into a tight bundle to introduce them into the brain from a single-entry point. The individual CFETs splay laterally in deep brain tissue during insertion due to their innate flexibility. This spatial redistribution allows navigation of the CFETs towards deep brain targets spreading horizontally from the axis of insertion. Commercial "linear" arrays provide single-entry insertion but only allow measurements along the axis of insertion. Horizontally configured arrays inflict separate penetrations for each individual channel. We tested functional performance of our CFET arrays in vivo for recording dopamine and for providing lateral spread to multiple distributed sites in the rat striatum. Spatial spread was further characterized in agar brain phantoms as a function of insertion depth. We also developed protocols to slice the embedded CFETs within fixed brain tissue using standard histology. This method allowed extraction of the precise spatial coordinates of the implanted CFETs and their recording sites as integrated with immunohistochemical staining for surrounding anatomical, cytological, and protein expression labels. Our CFET array has the potential to unlock a wide range of applications, from uncovering the role of neuromodulators in synaptic plasticity, to addressing critical safety barriers in clinical translation towards diagnostic and adaptive treatment in Parkinson's disease and major mood disorders.

Scalable, flexible carbon fiber electrode thread arrays for three-dimensional spatial profiling of neurochemical activity in deep brain structures of rodents.

We developed a flexible "electrode-thread" array for recording dopamine neurochemical activity from a lateral distribution of subcortical targets (up to 16) transverse to the axis of insertion. Ultrathin (∼ 10 µm diameter) carbon fiber (CF) electrode-threads (CFETs) are clustered into a tight bundle to introduce them into the brain from a single entry point. The individual CFETs splay laterally in deep brain tissue during insertion due to their innate flexibility. This spatial redistribution allows navigation of the CFETs towards deep brain targets spreading horizontally from the axis of insertion. Commercial "linear" arrays provide single entry insertion but only allow measurements along the axis of insertion. Horizontally configured neurochemical recording arrays inflict separate penetrations for each individual channel (i.e., electrode). We tested functional performance of our CFET arrays in vivo for recording dopamine neurochemical dynamics and for providing lateral spread to multiple distributed sites in the striatum of rats. Spatial spread was further characterized using agar brain phantoms to measure electrode deflection as a function of insertion depth. We also developed protocols to slice the embedded CFETs within fixed brain tissue using standard histology techniques. This method allowed extraction of the precise spatial coordinates of the implanted CFETs and their recording sites as integrated with immunohistochemical staining for surrounding anatomical, cytological, and protein expression labels. Neurochemical recording operations tested here can be integrated with already widely established capabilities of CF-based electrodes to record single neuron activity and local field potentials, to enable multi-modal recording functions. Our CFET array has the potential to unlock a wide range of applications, from uncovering the role of neuromodulators in synaptic plasticity, to addressing critical safety barriers in clinical translation towards diagnostic and adaptive treatment in Parkinson's disease and major mood disorders.

Synchronous Measurements of Extracellular Action Potentials and Neurochemical Activity with Carbon Fiber Electrodes in Nonhuman Primates.

Measuring the dynamic relationship between neuromodulators, such as dopamine, and neuronal action potentials is imperative to understand how these fundamental modes of neural signaling interact to mediate behavior. Here, we developed methods to measure concurrently dopamine and extracellular action potentials (i.e., spikes) and applied these in a monkey performing a behavioral task. Standard fast-scan cyclic voltammetric (FSCV) electrochemical (EChem) and electrophysiological (EPhys) recording systems are combined and used to collect spike and dopamine signals, respectively, from an array of carbon fiber (CF) sensors implanted in the monkey striatum. FSCV requires the application of small voltages at the implanted sensors to measure redox currents generated from target molecules, such as dopamine. These applied voltages create artifacts at neighboring EPhys-measurement sensors, producing signals that may falsely be classified as physiological spikes. Therefore, simple automated temporal interpolation algorithms were designed to remove these artifacts and enable accurate spike extraction. We validated these methods using simulated artifacts and demonstrated an average spike recovery rate of 84.5%. This spike extraction was performed on data collected from concurrent EChem and EPhys recordings made in a task-performing monkey to discriminate cell-type specific striatal units. These identified units were shown to correlate to specific behavioral task parameters related to reward size and eye-movement direction. Synchronous measures of spike and dopamine signals displayed contrasting relations to the behavioral task parameters, as taken from our small set of representative data, suggesting a complex relationship between these two modes of neural signaling. Future application of our methods will help advance our understanding of the interactions between neuromodulator signaling and neuronal activity, to elucidate more detailed mechanisms of neural circuitry and plasticity mediating behaviors in health and in disease.

Cellular-scale probes enable stable chronic subsecond monitoring of dopamine neurochemicals in a rodent model.

Chemical signaling underlies both temporally phasic and extended activity in the brain. Phasic activity can be monitored by implanted sensors, but chronic recording of such chemical signals has been difficult because the capacity to measure them degrades over time. This degradation has been attributed to tissue damage progressively produced by the sensors and failure of the sensors themselves. We report methods that surmount these problems through the development of sensors having diameters as small as individual neuronal cell bodies (<10 µm). These micro-invasive probes (µIPs) markedly reduced expression of detectable markers of inflammation and tissue damage in a rodent test model. The chronically implanted µIPs provided stable operation in monitoring sub-second fluctuations in stimulation-evoked dopamine in anesthetized rats for over a year. These findings demonstrate that monitoring of chemical activity patterns in the brain over at least year-long periods, long a goal of both basic and clinical neuroscience, is achievable.

Miniaturized neural system for chronic, local intracerebral drug delivery.

Recent advances in medications for neurodegenerative disorders are expanding opportunities for improving the debilitating symptoms suffered by patients. Existing pharmacologic treatments, however, often rely on systemic drug administration, which result in broad drug distribution and consequent increased risk for toxicity. Given that many key neural circuitries have sub-cubic millimeter volumes and cell-specific characteristics, small-volume drug administration into affected brain areas with minimal diffusion and leakage is essential. We report the development of an implantable, remotely controllable, miniaturized neural drug delivery system permitting dynamic adjustment of therapy with pinpoint spatial accuracy. We demonstrate that this device can chemically modulate local neuronal activity in small (rodent) and large (nonhuman primate) animal models, while simultaneously allowing the recording of neural activity to enable feedback control.

Long-term dopamine neurochemical monitoring in primates.

Many debilitating neuropsychiatric and neurodegenerative disorders are characterized by dopamine neurotransmitter dysregulation. Monitoring subsecond dopamine release accurately and for extended, clinically relevant timescales is a critical unmet need. Especially valuable has been the development of electrochemical fast-scan cyclic voltammetry implementing microsized carbon fiber probe implants to record fast millisecond changes in dopamine concentrations. Nevertheless, these well-established methods have only been applied in primates with acutely (few hours) implanted sensors. Neurochemical monitoring for long timescales is necessary to improve diagnostic and therapeutic procedures for a wide range of neurological disorders. Strategies for the chronic use of such sensors have recently been established successfully in rodents, but new infrastructures are needed to enable these strategies in primates. Here we report an integrated neurochemical recording platform for monitoring dopamine release from sensors chronically implanted in deep brain structures of nonhuman primates for over 100 days, together with results for behavior-related and stimulation-induced dopamine release. From these chronically implanted probes, we measured dopamine release from multiple sites in the striatum as induced by behavioral performance and reward-related stimuli, by direct stimulation, and by drug administration. We further developed algorithms to automate detection of dopamine. These algorithms could be used to track the effects of drugs on endogenous dopamine neurotransmission, as well as to evaluate the long-term performance of the chronically implanted sensors. Our chronic measurements demonstrate the feasibility of measuring subsecond dopamine release from deep brain circuits of awake, behaving primates in a longitudinally reproducible manner.

Subcellular probes for neurochemical recording from multiple brain sites.

Dysregulation of neurochemicals, in particular, dopamine, is epitomized in numerous debilitating disorders that impair normal movement and mood aspects of our everyday behavior. Neurochemical transmission is a neuron-specific process, and further exhibits region-specific signaling in the brain. Tools are needed to monitor the heterogeneous spatiotemporal dynamics of dopamine neurotransmission without compromising the physiological processes of the neuronal environment. We developed neurochemical probes that are ten times smaller than any existing dopamine sensor, based on the size of the entire implanted shaft and its sensing tip. The microfabricated probe occupies a spatial footprint (9 µm) coordinate with the average size of individual neuronal cells (∼10 µm). These cellular-scale probes were shown to reduce inflammatory response of the implanted brain tissue environment. The probes are further configured in the form of a microarray to permit electrochemical sampling of dopamine and other neurotransmitters at unprecedented spatial densities and distributions. Dopamine recording was performed concurrently from up to 16 sites in the striatum of rats, revealing a remarkable spatiotemporal contrast in dopamine transmission as well as site-specific pharmacological modulation. Collectively, the reported platform endeavors to enable high density mapping of the chemical messengers fundamentally involved in neuronal communication through the use of minimally invasive probes that help preserve the neuronal viability of the implant environment.

In vitro hydrodynamic, transient, and overtime performance of a miniaturized valve for hydrocephalus.

Reliable cerebrospinal fluid (CSF) draining methods are needed to treat hydrocephalus, a chronic debilitating brain disorder. Current shunt implant treatments are characterized by high failure rates that are to some extent attributed to their length and multiple components. The designed valve, made of hydrogel, steers away from such protracted schemes and intends to provide a direct substitute for faulty arachnoid granulations, the brain's natural CSF draining valves, and restore CSF draining operations within the cranium. The valve relies on innate hydrogel swelling phenomena to strengthen reverse flow sealing at idle and negative pressures thereby alleviating common valve failure mechanisms. In vitro measurements display operation in range of natural CSF draining (cracking pressure, PT ~ 1-110 mmH2O and outflow hydraulic resistance, Rh ~ 24-152 mmH2O/mL/min), with negligible reverse flow leakage (flow, QO > -10 µL/min). Hydrodynamic measurements and over-time tests under physically relevant conditions further demonstrate the valve's operationally-reproducible properties and strengthen its validity for use as a chronic implant.

Miniaturized passive hydrogel check valve for hydrocephalus treatment.

Improvements in cerebrospinal fluid (CSF) draining techniques for treatment of hydrocephalus are urgently sought after to substitute for current CSF shunts that are plagued by high failure rates. The passive check valve aims to restore near natural CSF draining operations while mitigating possible failure mechanisms caused by finite leakage or low resilience that frequently constrain practical implementation of miniaturized valves. A simple hydrogel diaphragm structures core passive valve operations and enforce valve sealing properties to substantially lower reverse flow leakage. Experimental measurements demonstrate realization of targeted cracking pressures (PT ≈ 20-110 mmH2O) and operation at -800 <; ΔP <; 600 mmH2O without observable degradation or leakage.

A Fully-Passive Wireless Microsystem for Recording of Neuropotentials using RF Backscattering Methods.

The ability to safely monitor neuropotentials is essential in establishing methods to study the brain. Current research focuses on the wireless telemetry aspect of implantable sensors in order to make these devices ubiquitous and safe. Chronic implants necessitate superior reliability and durability of the integrated electronics. The power consumption of implanted electronics must also be limited to within several milliwatts to microwatts to minimize heat trauma in the human body. In order to address these severe requirements, we developed an entirely passive and wireless microsystem for recording neuropotentials. An external interrogator supplies a fundamental microwave carrier to the microsystem. The microsystem comprises varactors that perform nonlinear mixing of neuropotential and fundamental carrier signals. The varactors generate third-order mixing products that are wirelessly backscattered to the external interrogator where the original neuropotential signals are recovered. Performance of the neuro-recording microsystem was demonstrated by wireless recording of emulated and in vivo neuropotentials. The obtained results were wireless recovery of neuropotentials as low as approximately 500 microvolts peak-to-peak (µV(pp)) with a bandwidth of 10 Hz to 3 kHz (for emulated signals) and with 128 epoch signal averaging of repetitive signals (for in vivo signals).