Membrane depolarization mediates both the inhibition of neural activity and cell-type-differences in response to high-frequency stimulation.

Neuromodulation using high frequency (>1 kHz) electric stimulation (HFS) enables preferential activation or inhibition of individual neural types, offering the possibility of more effective treatments across a broad spectrum of neurological diseases. To improve effectiveness, it is important to better understand the mechanisms governing activation and inhibition with HFS so that selectivity can be optimized. In this study, we measure the membrane potential (Vm) and spiking responses of ON and OFF α-sustained retinal ganglion cells (RGCs) to a wide range of stimulus frequencies (100-2500 Hz) and amplitudes (10-100 µA). Our findings indicate that HFS induces shifts in Vm, with both the strength and polarity of the shifts dependent on the stimulus conditions. Spiking responses in each cell directly correlate with the shifts in Vm, where strong depolarization leads to spiking suppression. Comparisons between the two cell types reveal that ON cells are more depolarized by a given amplitude of HFS than OFF cells-this sensitivity difference enables the selective targeting. Computational modeling indicates that ion-channel dynamics largely account for the shifts in Vm, suggesting that a better understanding of the differences in ion-channel properties across cell types may improve the selectivity and ultimately, enhance HFS-based neurostimulation strategies.

Flexible, scalable, high channel count stereo-electrode for recording in the human brain.

Over the past decade, stereotactically placed electrodes have become the gold standard for deep brain recording and stimulation for a wide variety of neurological and psychiatric diseases. Current electrodes, however, are limited in their spatial resolution and ability to record from small populations of neurons, let alone individual neurons. Here, we report on an innovative, customizable, monolithically integrated human-grade flexible depth electrode capable of recording from up to 128 channels and able to record at a depth of 10 cm in brain tissue. This thin, stylet-guided depth electrode is capable of recording local field potentials and single unit neuronal activity (action potentials), validated across species. This device represents an advance in manufacturing and design approaches which extends the capabilities of a mainstay technology in clinical neurology.

Thermal safety considerations for implantable micro-coil design.

Micro magnetic stimulation of the brain via implantable micro-coils is a promising novel technology for neuromodulation. Careful consideration of the thermodynamic profile of such devices is necessary for effective and safe designs.Objective.We seek to quantify the thermal profile of bent wire micro-coils in order to understand and mitigate thermal impacts of micro-coil stimulation.Approach. In this study, we use fine wire thermocouples and COMSOL finite element modeling to examine the profile of the thermal gradients generated near bent wire micro-coils submerged in a water bath during stimulation. We tested a range of stimulation parameters previously reported in the literature such as voltage amplitude, stimulus frequency, stimulus repetition rate and coil wire materials.Main results. We found temperature increases ranging from <1 °C to 8.4 °C depending upon the stimulation parameters tested and coil wire materials used. Numerical modeling of the thermodynamics identified hot spots of the highest temperatures along the micro-coil contributing to the thermal gradients and demonstrated that these thermal gradients can be mitigated by the choice of wire conductor material and construction geometry.Significance. ISO standard 14708-1 designates a thermal safety limit of 2 °C temperature increase for active implantable medical devices. By switching the coil wire material from platinum/iridium to gold, our study achieved a 5-6-fold decrease in the thermal impact of coil stimulation. The thermal gradients generated from the gold wire coil were measured below the 2 °C safety limit for all stimulation parameters tested.

Layer-dependent stability of intracortical recordings and neuronal cell loss.

Intracortical recordings can be used to voluntarily control external devices via brain-machine interfaces (BMI). Multiple factors, including the foreign body response (FBR), limit the stability of these neural signals over time. Current clinically approved devices consist of multi-electrode arrays with a single electrode site at the tip of each shank, confining the recording interface to a single layer of the cortex. Advancements in manufacturing technology have led to the development of high-density electrodes that can record from multiple layers. However, the long-term stability of neural recordings and the extent of neuronal cell loss around the electrode across different cortical depths have yet to be explored. To answer these questions, we recorded neural signals from rats chronically implanted with a silicon-substrate microelectrode array spanning the layers of the cortex. Our results show the long-term stability of intracortical recordings varies across cortical depth, with electrode sites around L4-L5 having the highest stability. Using machine learning guided segmentation, our novel histological technique, DeepHisto, revealed that the extent of neuronal cell loss varies across cortical layers, with L2/3 and L4 electrodes having the largest area of neuronal cell loss. These findings suggest that interfacing depth plays a major role in the FBR and long-term performance of intracortical neuroprostheses.

MEMS micro-coils for magnetic neurostimulation.

Micro-coil magnetic stimulation of brain tissue presents new challenges for MEMS micro-coil probe fabrication. The main challenges are threefold; (i) low coil resistance for high power efficiency, (ii) low leak current from the probe into the in vitro experimental set-up, (iii) adaptive MEMS process technology because of the dynamic research area, which requires agile design changes. Taking on these challenges, we present a MEMS fabrication process that has three main features; (i) multilayer resist lift-off process to pattern up to 1800-nm-thick metal films, and special care is taken to obtain high conductivity thin-films by physical vapor deposition, and (ii) all micro-coil Al wires are encapsulated in at least 200 nm of ALD alumina and 6-µm-thick parylene C such the leak resistance is high (>210 GΩ), (iii) combining a multi-step DRIE process and maskless photolithography for adaptive design and device fabrication. The entire process requires four lithography steps. Because we avoided SOI wafers and lithography mask fabrication, the design-to-device time is shortened significantly. The resulting probes are 4-mm-long, 60-µm-thick, and down to 150 µm-wide. Selected MEMS coil devices were validated in vivo using mice and compared to previous work.

Thermal effects on neurons during stimulation of the brain.

All electric and magnetic stimulation of the brain deposits thermal energy in the brain. This occurs through either Joule heating of the conductors carrying current through electrodes and magnetic coils, or through dissipation of energy in the conductive brain.Objective.Although electrical interaction with brain tissue is inseparable from thermal effects when electrodes are used, magnetic induction enables us to separate Joule heating from induction effects by contrasting AC and DC driving of magnetic coils using the same energy deposition within the conductors. Since mammalian cortical neurons have no known sensitivity to static magnetic fields, and if there is no evidence of effect on spike timing to oscillating magnetic fields, we can presume that the induced electrical currents within the brain are below the molecular shot noise where any interaction with tissue is purely thermal.Approach.In this study, we examined a range of frequencies produced from micromagnetic coils operating below the molecular shot noise threshold for electrical interaction with single neurons.Main results.We found that small temperature increases and decreases of 1∘C caused consistent transient suppression and excitation of neurons during temperature change. Numerical modeling of the biophysics demonstrated that the Na-K pump, and to a lesser extent the Nernst potential, could account for these transient effects. Such effects are dependent upon compartmental ion fluxes and the rate of temperature change.Significance.A new bifurcation is described in the model dynamics that accounts for the transient suppression and excitation; in addition, we note the remarkable similarity of this bifurcation's rate dependency with other thermal rate-dependent tipping points in planetary warming dynamics. These experimental and theoretical findings demonstrate that stimulation of the brain must take into account small thermal effects that are ubiquitously present in electrical and magnetic stimulation. More sophisticated models of electrical current interaction with neurons combined with thermal effects will lead to more accurate modulation of neuronal activity.

Micro-magnetic stimulation of primary visual cortex induces focal and sustained activation of secondary visual cortex.

Cortical visual prostheses that aim to restore sight to the blind require the ability to create neural activity in the visual cortex. Electric stimulation delivered via microelectrodes implanted in the primary visual cortex (V1) has been the most common approach, although conventional electrodes may not effectively confine activation to focal regions and thus the acuity they create may be limited. Magnetic stimulation from microcoils confines activation to single cortical columns of V1 and thus may prove to be more effective than conventional microelectrodes, but the ability of microcoils to drive synaptic connections has not been explored. Here, we show that magnetic stimulation of V1 using microcoils induces spatially confined activation in the secondary visual cortex (V2) in mouse brain slices. Single-loop microcoils were fabricated using platinum-iridium flat microwires, and their effectiveness was evaluated using calcium imaging and compared with that of monopolar and bipolar electrodes. Our results show that compared to the electrodes, the microcoils better confined activation to a small region in V1. In addition, they produced more precise and sustained activation in V2. The finding that microcoil-based stimulation propagates to higher visual centres raises the possibility that complex visual perception, e.g. that requiring sustained synaptic inputs, may be achievable. This article is part of the theme issue 'Advanced neurotechnologies: translating innovation for health and well-being'.

The Long-Term Stability of Intracortical Microstimulation and the Foreign Body Response Are Layer Dependent.

Intracortical microstimulation (ICMS) of the somatosensory cortex (S1) can restore sensory function in patients with paralysis. Studies assessing the stability of ICMS have reported heterogeneous responses across electrodes and over time, potentially hindering the implementation and translatability of these technologies. The foreign body response (FBR) and the encapsulating glial scar have been associated with a decay in chronic performance of implanted electrodes. Moreover, the morphology, intrinsic properties, and function of cells vary across cortical layers, each potentially affecting the sensitivity to ICMS as well as the degree of the FBR across cortical depth. However, layer-by-layer comparisons of the long-term stability of ICMS as well as the extent of the astrocytic glial scar change across cortical layers have not been well explored. Here, we implanted silicon microelectrodes with electrode sites spanning all the layers of S1 in rats. Using a behavioral paradigm, we obtained ICMS detection thresholds from all cortical layers for up to 40 weeks. Our results showed that the sensitivity and long-term performance of ICMS is indeed layer dependent. Overall, detection thresholds decreased during the first 7 weeks post-implantation (WPI). This was followed by a period in which thresholds remained stable or increased depending on the interfacing layer: thresholds in L1 and L6 exhibited the most consistent increases over time, while those in L4 and L5 remained the most stable. Furthermore, histological investigation of the tissue surrounding the electrode showed a biological response of microglia and macrophages which peaked at L1, while the area of the astrocytic glial scar peaked at L2/3. Interestingly, the biological response of these FBR markers is less exacerbated at L4 and L5, suggesting a potential link between the FBR and the long-term stability of ICMS. These findings suggest that interfacing depth can play an important role in the design of chronically stable implantable microelectrodes.

Spiking Characteristics of Network-Mediated Responses Arising in Direction-Selective Ganglion Cells of Rabbit and Mouse Retinas to Electric Stimulation for Retinal Prostheses.

To restore the sight of individuals blinded by outer retinal degeneration, numerous retinal prostheses have been developed. However, the performance of those implants is still hampered by some factors including the lack of comprehensive understanding of the electrically-evoked responses arising in various retinal ganglion cell (RGC) types. In this study, we characterized the electrically-evoked network-mediated responses (hereafter referred to as electric responses) of ON-OFF direction-selective (DS) RGCs in rabbit and mouse retinas for the first time. Interestingly, both species in common demonstrated strong negative correlations between spike counts of electric responses and direction selective indices (DSIs), suggesting electric stimulation activates inhibitory presynaptic neurons that suppress null direction responses for high direction tuning in their light responses. The DS cells of the two species showed several differences including different numbers of bursts. Also, spiking patterns were more heterogeneous across DS RGCs of rabbits than those of mice. The electric response magnitudes of rabbit DS cells showed positive and negative correlations with ON and OFF light response magnitudes to preferred direction motion, respectively. But the mouse DS cells showed positive correlations in both comparisons. Our Fano Factor (FF) and spike time tiling coefficient (STTC) analyses revealed that spiking consistencies across repeats were reduced in late electric responses in both species. Moreover, the response consistencies of DS RGCs were lower than those of non-DS RGCs. Our results indicate the species-dependent retinal circuits may result in different electric response features and therefore suggest a proper animal model may be crucial in prosthetic researches.

MEMS inductor fabrication and emerging applications in power electronics and neurotechnologies.

MEMS inductors are used in a wide range of applications in micro- and nanotechnology, including RF MEMS, sensors, power electronics, and Bio-MEMS. Fabrication technologies set the boundary conditions for inductor design and their electrical and mechanical performance. This review provides a comprehensive overview of state-of-the-art MEMS technologies for inductor fabrication, presents recent advances in 3D additive fabrication technologies, and discusses the challenges and opportunities of MEMS inductors for two emerging applications, namely, integrated power electronics and neurotechnologies. Among the four top-down MEMS fabrication approaches, 3D surface micromachining and through-substrate-via (TSV) fabrication technology have been intensively studied to fabricate 3D inductors such as solenoid and toroid in-substrate TSV inductors. While 3D inductors are preferred for their high-quality factor, high power density, and low parasitic capacitance, in-substrate TSV inductors offer an additional unique advantage for 3D system integration and efficient thermal dissipation. These features make in-substrate TSV inductors promising to achieve the ultimate goal of monolithically integrated power converters. From another perspective, 3D bottom-up additive techniques such as ice lithography have great potential for fabricating inductors with geometries and specifications that are very challenging to achieve with established MEMS technologies. Finally, we discuss inspiring and emerging research opportunities for MEMS inductors.

The impact of synchronous versus asynchronous electrical stimulation in artificial vision.

Visual prosthesis devices designed to restore sight to the blind have been under development in the laboratory for several decades. Clinical translation continues to be challenging, due in part to gaps in our understanding of critical parameters such as how phosphenes, the electrically-generated pixels of artificial vision, can be combined to form images. In this review we explore the effects that synchronous and asynchronous electrical stimulation across multiple electrodes have in evoking phosphenes. Understanding how electrical patterns influence phosphene generation to control object binding and perception of visual form is fundamental to creation of a clinically successful prosthesis.

Microscale Physiological Events on the Human Cortical Surface.

Despite ongoing advances in our understanding of local single-cellular and network-level activity of neuronal populations in the human brain, extraordinarily little is known about their "intermediate" microscale local circuit dynamics. Here, we utilized ultra-high-density microelectrode arrays and a rare opportunity to perform intracranial recordings across multiple cortical areas in human participants to discover three distinct classes of cortical activity that are not locked to ongoing natural brain rhythmic activity. The first included fast waveforms similar to extracellular single-unit activity. The other two types were discrete events with slower waveform dynamics and were found preferentially in upper cortical layers. These second and third types were also observed in rodents, nonhuman primates, and semi-chronic recordings from humans via laminar and Utah array microelectrodes. The rates of all three events were selectively modulated by auditory and electrical stimuli, pharmacological manipulation, and cold saline application and had small causal co-occurrences. These results suggest that the proper combination of high-resolution microelectrodes and analytic techniques can capture neuronal dynamics that lay between somatic action potentials and aggregate population activity. Understanding intermediate microscale dynamics in relation to single-cell and network dynamics may reveal important details about activity in the full cortical circuit.

Layer-specific parameters of intracortical microstimulation of the somatosensory cortex.

Objective. Intracortical microstimulation of the primary somatosensory cortex (S1) has shown great progress in restoring touch sensations to patients with paralysis. Stimulation parameters such as amplitude, phase duration, and frequency can influence the quality of the evoked percept as well as the amount of charge necessary to elicit a response. Previous studies in V1 and auditory cortices have shown that the behavioral responses to stimulation amplitude and phase duration change across cortical depth. However, this depth-dependent response has yet to be investigated in S1. Similarly, to our knowledge, the response to microstimulation frequency across cortical depth remains unexplored.Approach. To assess these questions, we implanted rats in S1 with a microelectrode with electrode-sites spanning all layers of the cortex. A conditioned avoidance behavioral paradigm was used to measure detection thresholds and responses to phase duration and frequency across cortical depth.Main results. Analogous to other cortical areas, the sensitivity to charge and strength-duration chronaxies in S1 varied across cortical layers. Likewise, the sensitivity to microstimulation frequency was layer dependent.Significance. These findings suggest that cortical depth can play an important role in the fine-tuning of stimulation parameters and in the design of intracortical neuroprostheses for clinical applications.

Morphological Factors that Underlie Neural Sensitivity to Stimulation in the Retina.

Retinal prostheses are a promising therapeutic intervention for patients afflicted by outer retinal degenerative diseases like retinitis pigmentosa and age-related macular degeneration. While significant advances in the development of retinal implants have been made, the quality of vision elicited by these devices remains largely sub-optimal. The variability in the responses produced by retinal devices is most likely due to the differences between the natural cell type-specific signaling that occur in the healthy retina vs. the non-specific activation of multiple cell types arising from artificial stimulation. In order to replicate these natural signaling patterns, stimulation strategies must be capable of preferentially activating specific RGC types. To design more selective stimulation strategies, a better understanding of the morphological factors that underlie the sensitivity to prosthetic stimulation must be developed. This review will focus on the role that different anatomical components play in driving the direct activation of RGCs by extracellular stimulation. Briefly, it will (1) characterize the variability in morphological properties of α-RGCs, (2) detail the influence of morphology on the direct activation of RGCs by electric stimulation, and (3) describe some of the potential biophysical mechanisms that could explain differences in activation thresholds and electrically evoked responses between RGC types.

Differential Responses to High-Frequency Electrical Stimulation in Brisk-Transient and Delta Retinal Ganglion Cells.

Retinal microprostheses strive to evoke a sense of vision in individuals blinded by outer retinal degenerative diseases, by electrically stimulating the surviving retina. It is widely suspected that a stimulation strategy that can selectively activate different retinal ganglion cell types will improve the quality of evoked phosphenes. Previous efforts towards this goal demonstrated the potential for selective ON and OFF brisk-transient cell activation using high-rate (2000 pulses per second, PPS) stimulation. Here, we build upon this earlier work by testing an additional rate of stimulation and additional cell populations. We find considerable variability in responses both within and across individual cell types, but show that the sensitivity of a ganglion cell to repetitive stimulation is highly correlated to its single-pulse threshold. Consistent with this, we found thresholds for both stimuli to be correlated to soma size, and thus likely mediated by the properties of the axon initial segment. The ultimate efficacy of high-rate stimulation will likely depend on several factors, chief among which are (a) the residual ganglion types, and (b) the stimulation frequency.

Response Profiles of Retinal Ganglion Cells to Sinusoidal Electric Stimulation vary for Low vs. High Frequencies.

Microelectronic retinal prostheses electrically stimulate retinal neurons with the goal of restoring vision in patients blinded by outer retinal degeneration. Despite some success in clinical trials, the quality of vision elicited by these devices is still limited. To improve the performance of retinal prostheses, our group studied how retinal neurons respond to electric stimulation. Our previous work showed that responses of retinal ganglion cells (RGCs) are frequency-dependent and different types of RGCs can be preferentially activated with a specific frequency and current amplitude. In the present study, we systemically examined responses of RGCs to sinusoidal electric stimulation with varying frequencies and amplitudes. We found that ON sustained alpha RGCs show distinct stimulus-response relationships to low and high frequency stimulation. For example, RGCs showed monotonic response curves to 500 Hz sinusoidal stimulation, whereas they showed non-monotonic response curves to 2000 Hz stimulation. We also described how increasing stimulus frequency gradually changed the response curves of RGCs.

Tailoring of the axon initial segment shapes the conversion of synaptic inputs into spiking output in OFF-α T retinal ganglion cells.

Recently, mouse OFF-α transient (OFF-α T) retinal ganglion cells (RGCs) were shown to display a gradient of light responses as a function of position along the dorsal-ventral axis; response differences were correlated to differences in the level of excitatory presynaptic input. Here, we show that postsynaptic differences between cells also make a strong contribution to response differences. Cells in the dorsal retina had longer axon initial segments (AISs)-the greater number of Nav1.6 channels in longer AISs directly mediates higher rates of spiking and helps avoid depolarization block that terminates spiking in ventral cells with shorter AISs. The pre- and postsynaptic specializations that shape the output of OFF-α T RGCs interact in different ways: In dorsal cells, strong inputs and the long AISs are both necessary to generate their strong, sustained spiking outputs, while in ventral cells, weak inputs or the short AISs are both sufficient to limit the spiking signal.

Spatially confined responses of mouse visual cortex to intracortical magnetic stimulation from micro-coils.

OBJECTIVE: Electrical stimulation via microelectrodes implanted in cortex has been suggested as a potential treatment for a wide range of neurological disorders. Despite some success however, the effectiveness of conventional electrodes remains limited, in part due to an inability to create specific patterns of neural activity around each electrode and in part due to challenges with maintaining a stable interface. The use of implantable micro-coils to magnetically stimulate the cortex has the potential to overcome these limitations because the asymmetric fields from coils can be harnessed to selectively activate some neurons, e.g. vertically-oriented pyramidal neurons while avoiding others, e.g. horizontally-oriented passing axons. In vitro experiments have shown that activation is indeed confined with micro-coils but their effectiveness in the intact brain of living animals has not been evaluated. APPROACH: To assess the efficacy of stimulation, a 128-channel custom recording microelectrode array was positioned on the surface of the visual cortex (ECoG) in anesthetized mice and responses to magnetic and electric stimulation were compared. Stimulation was delivered from electrodes or micro-coils implanted through a hole in the center of the recording array at a rate of 200 pulses per second for 100 ms. MAIN RESULTS: Both electric and magnetic stimulation reliably elicited cortical responses, although activation from electric stimulation was spatially expansive, often extending more than 1 mm from the stimulation site, while activation from magnetic stimulation was typically confined to a ∼300 µm diameter region around the stimulation site. Results were consistent for stimulation of both cortical layer 2/3 and layer 5 as well as across a range of stimulus strengths. SIGNIFICANCE: The improved focality with magnetic stimulation suggests that the effectiveness of cortical stimulation can be improved. Improved focality may be particularly attractive for cortical prostheses that require high spatial resolution, e.g. devices that target sensory cortex, as it may lead to improved acuity.

Retinal Degeneration Reduces Consistency of Network-Mediated Responses Arising in Ganglion Cells to Electric Stimulation.

Retinal prostheses use periodic repetition of electrical stimuli to form artificial vision. To enhance the reliability of evoked visual percepts, repeating stimuli need to evoke consistent spiking activity in individual retinal ganglion cells (RGCs). However, it is not well known whether outer retinal degeneration alters the consistency of RGC responses. Hence, here we systematically investigated the trial-to-trial variability in network-mediated responses as a function of the degeneration level. We patch-clamp recorded spikes in ON and OFF types of alpha RGCs from r d10 mice at four different postnatal days (P15, P19, P31, and P60), representing distinct stages of degeneration. To assess the consistency of responses, we analyzed variances in spike count and timing across repeats of the same stimulus delivered multiple times. We found the trial-to-trial variability of network-mediated responses increased considerably as the disease progressed. Compared to responses taken before degeneration onset, those of degenerate retinas showed up to ~70% higher variability (Fano Factor) in spike counts (p < 0.001) and ~95% lower correlation level in spike timing (p < 0.001). These results indicate consistency weakens significantly in electrically-evoked network-mediated responses and therefore raise concerns about the ability of microelectronic retinal implants to elicit consistent visual percepts at advanced stages of retinal degeneration.