Recordings In Non-human Primates Research Articles

A high-density 1,024-channel probe for brain-wide recordings in non-human primates.

Large-scale neural population recordings with single-cell resolution across the primate brain remain challenging. Here we introduce the Neuroscroll probe that isolates single neuronal activities simultaneously from 1,024 densely spaced channels that are flexibly distributed across the shank of the probe. The Neuroscroll probe length is easily tunable for individual probes from 10 mm to 90 mm, covering the brain size of non-human primates and humans, and the probes remain intact and functional after repeated bending deformations. The Neuroscroll probes provided reliable recordings from large neural populations with high chronic stability up to 105 weeks in rats. Recording with each Neuroscroll probe yielded hundreds of well-isolated single units simultaneously from multiple brain regions distributed across the entire depth of the rhesus macaque brain. With the thousand simultaneously recorded channels, unprecedented probe length, excellent mechanical stability and flexible recording site distribution, the Neuroscroll probes enable a wide range of new experimental paradigms in system neuroscience studies with great versatility.

Temporal interference stimulation disrupts spike timing in the primate brain

Electrical stimulation can regulate brain activity, producing clear clinical benefits, but focal and effective neuromodulation often requires surgically implanted electrodes. Recent studies argue that temporal interference (TI) stimulation may provide similar outcomes non-invasively. During TI, scalp electrodes generate multiple electrical fields in the brain, modulating neural activity only at their intersection. Despite considerable enthusiasm for this approach, little empirical evidence demonstrates its effectiveness, especially under conditions suitable for human use. Here, using single-neuron recordings in non-human primates, we establish that TI reliably alters the timing, but not the rate, of spiking activity. However, we show that TI requires strategies—high carrier frequencies, multiple electrodes, and amplitude-modulated waveforms—that also limit its effectiveness. Combined, these factors make TI 80 % weaker than other forms of non-invasive brain stimulation. Although unlikely to cause widespread neuronal entrainment, TI may be ideal for disrupting pathological oscillatory activity, a hallmark of many neurological disorders.

Neuronal Modeling of Cross-Sensory Visual Evoked Magnetoencephalography Responses in the Auditory Cortex.

Previous studies have demonstrated that auditory cortex activity can be influenced by cross-sensory visual inputs. Intracortical laminar recordings in nonhuman primates have suggested a feedforward (FF) type profile for auditory evoked but feedback (FB) type for visual evoked activity in the auditory cortex. To test whether cross-sensory visual evoked activity in the auditory cortex is associated with FB inputs also in humans, we analyzed magnetoencephalography (MEG) responses from eight human subjects (six females) evoked by simple auditory or visual stimuli. In the estimated MEG source waveforms for auditory cortex regions of interest, auditory evoked response showed peaks at 37 and 90 ms and visual evoked response at 125 ms. The inputs to the auditory cortex were modeled through FF- and FB-type connections targeting different cortical layers using the Human Neocortical Neurosolver (HNN), which links cellular- and circuit-level mechanisms to MEG signals. HNN modeling suggested that the experimentally observed auditory response could be explained by an FF input followed by an FB input, whereas the cross-sensory visual response could be adequately explained by just an FB input. Thus, the combined MEG and HNN results support the hypothesis that cross-sensory visual input in the auditory cortex is of FB type. The results also illustrate how the dynamic patterns of the estimated MEG source activity can provide information about the characteristics of the input into a cortical area in terms of the hierarchical organization among areas.

An Ultraflexible Electrode Array for Large-Scale Chronic Recording in the Nonhuman Primate Brain.

Single-unit (SU) recording in nonhuman primates (NHPs) is indispensible in the quest of how the brain works, yet electrodes currently used for the NHP brain are limited in signal longevity, stability, and spatial coverage. Using new structural materials, microfabrication, and penetration techniques, we develop a mechanically robust ultraflexible, 1 µm thin electrode array (MERF) that enables pial penetration and high-density, large-scale, and chronic recording of neurons along both vertical and horizontal cortical axes in the nonhuman primate brain. Recording from three monkeys yields 2,913 SUs from 1,065 functional recording channels (up to 240 days), with some SUs tracked for up to 2 months. Recording from the primary visual cortex (V1) reveals that neurons with similar orientation preferences for visual stimuli exhibited higher spike correlation. Furthermore, simultaneously recorded neurons in different cortical layers of the primary motor cortex (M1) show preferential firing for hand movements of different directions. Finally, it is shown that a linear decoder trained with neuronal spiking activity across M1 layers during monkey's hand movements can be used to achieve on-line control of cursor movement. Thus, the MERF electrode array offers a new tool for basic neuroscience studies and brain-machine interface (BMI) applications in the primate brain.

Flexible multichannel electrodes for acute recording in nonhuman primates

Flexible electrodes have demonstrated better biocompatibility than rigid electrodes in relieving tissue encapsulation and long-term recording. Nonhuman primates are closer to humans in their brains’ structural and functional properties, thus making them more suitable than rodents as animal models for potential clinical usage. However, the application of flexible electrodes on nonhuman primates has rarely been reported. In the present study, a flexible multichannel electrode array for nonhuman primates was developed and implemented for extracellular recording in behaving monkeys. To minimize the window of durotomy for reducing possible risks, a guide-tube-compatible implantation solution was designed to deliver the flexible electrodes through the dura into the cortex. The proposed structure for inserting flexible electrodes was characterized ex vivo and validated in vivo. Furthermore, acute recording of multichannel flexible electrodes for the primates was performed. The results showed that the flexible electrodes and implantation method used in this study meet the needs of extracellular recording in nonhuman primates. Task-related neuronal activities with a high signal-to-noise ratio of spikes demonstrated that our whole device is currently a minimally invasive and clinically viable approach for extracellular recording.

3D printed guide tube system for acute Neuropixels probe recordings in non-human primates

Objective. Neuropixels (NP) probes are a significant advance in electrophysiological recording technology that enable monitoring of hundreds of neurons in the brain simultaneously at different depths. Application of this technology has been predominately in rodents, however widespread use in non-human primates (NHPs) such as rhesus macaques has been limited. In this study we sought to overcome two overarching challenges that impede acute NP implantation in NHPs: (1) traditional microdrive systems that mount to cephalic chambers are commonly used to access cortical areas for microelectrode recordings but are not designed to accommodate NP probes, and (2) NHPs have thick dura mater and tissue growth within the cephalic chambers which poses a challenge for insertion of the extremely fragile NP probe. Approach. In this study we present a novel NP guide tube system that can be adapted to commercial microdrive systems and demonstrate an implant method using the NP guide tube system. This system was developed using a combination of CAD design, 3D printing, and small part machining. Software programs, 3D Slicer and SolidWorks were used to target cortical areas, approximate recording depths and locations, and for in-silico implant testing. Main results. We performed in vivo testing to validate our methodology, successfully implanting, explanting, and reimplanting NP probes. We collected stable neurophysiological recordings in the premotor cortex of a rhesus macaque at rest and during performance of a reaching task. Significance. In this study we demonstrate a robust Neuropixels implant system that allows multiple penetrations with the same NP probe and share design files that will facilitate the adoption of this powerful recording technology for NHP studies.

An improved system for simultaneous transcranial magnetic stimulation and single-unit recordings in non-human primates

An improved system for simultaneous transcranial magnetic stimulation and single-unit recordings in non-human primates

Cortical depth profiles of auditory and visual 7T functional MRI responses in human superior temporal areas.

Invasive neurophysiological studies in nonhuman primates have shown different laminar activation profiles to auditory vs. visual stimuli in auditory cortices and adjacent polymodal areas. Means to examine the underlying feedforward vs. feedback type influences noninvasively have been limited in humans. Here, using 1-mm isotropic resolution 3D echo-planar imaging at 7 T, we studied the intracortical depth profiles of functional magnetic resonance imaging (fMRI) blood oxygenation level dependent (BOLD) signals to brief auditory (noise bursts) and visual (checkerboard) stimuli. BOLD percent-signal-changes were estimated at 11 equally spaced intracortical depths, within regions-of-interest encompassing auditory (Heschl's gyrus, Heschl's sulcus, planum temporale, and posterior superior temporal gyrus) and polymodal (middle and posterior superior temporal sulcus) areas. Effects of differing BOLD signal strengths for auditory and visual stimuli were controlled via normalization and statistical modeling. The BOLD depth profile shapes, modeled with quadratic regression, were significantly different for auditory vs. visual stimuli in auditory cortices, but not in polymodal areas. The different depth profiles could reflect sensory-specific feedforward versus cross-sensory feedback influences, previously shown in laminar recordings in nonhuman primates. The results suggest that intracortical BOLD profiles can help distinguish between feedforward and feedback type influences in the human brain. Further experimental studies are still needed to clarify how underlying signal strength influences BOLD depth profiles under different stimulus conditions.

An improved system for simultaneous transcranial magnetic stimulation and single-unit recordings in non-human primates

An improved system for simultaneous transcranial magnetic stimulation and single-unit recordings in non-human primates

Working Memory: From Neural Activity to the Sentient Mind.

Working memory (WM) is the ability to maintain and manipulate information in the conscious mind over a timescale of seconds. This ability is thought to be maintained through the persistent discharges of neurons in a network of brain areas centered on the prefrontal cortex, as evidenced by neurophysiological recordings in nonhuman primates, though both the localization and the neural basis of WM has been a matter of debate in recent years. Neural correlates of WM are evident in species other than primates, including rodents and corvids. A specialized network of excitatory and inhibitory neurons, aided by neuromodulatory influences of dopamine, is critical for the maintenance of neuronal activity. Limitations in WM capacity and duration, as well as its enhancement during development, can be attributed to properties of neural activity and circuits. Changes in these factors can be observed through training-induced improvements and in pathological impairments. WM thus provides a prototypical cognitive function whose properties can be tied to the spiking activity of brain neurons. © 2021 American Physiological Society. Compr Physiol 11:1-41, 2021.

Pruriception and neuronal coding in nociceptor subtypes in human and nonhuman primates.

In humans, intradermal administration of β-alanine (ALA) and bovine adrenal medulla peptide 8-22 (BAM8-22) evokes the sensation of itch. Currently, it is unknown which human dorsal root ganglion (DRG) neurons express the receptors of these pruritogens, MRGPRD and MRGPRX1, respectively, and which cutaneous afferents these pruritogens activate in primate. In situ hybridization studies revealed that MRGPRD and MRGPRX1 are co-expressed in a subpopulation of TRPV1+ human DRG neurons. In electrophysiological recordings in nonhuman primates (Macaca nemestrina), subtypes of polymodal C-fiber nociceptors are preferentially activated by ALA and BAM8-22, with significant overlap. When pruritogens ALA, BAM8-22, and histamine, which activate different subclasses of C-fiber afferents, are administered in combination, human volunteers report itch and nociceptive sensations similar to those induced by a single pruritogen. Our results provide evidence for differences in pruriceptive processing between primates and rodents, and do not support the spatial contrast theory of coding of itch and pain.

Differential attention-dependent adjustment of frequency, power and phase in primary sensory and frontoparietal areas

Continuously prioritizing behaviourally relevant information from the environment for improved stimulus processing is a crucial function of attention. In the current MEG study, we investigated how ongoing oscillatory activity of both sensory and non-sensory brain regions are differentially impacted by attentional focus. Low-frequency phase alignment of neural activity in primary sensory areas, with respect to attended/ignored features has been suggested to support top-down prioritization. However, phase adjustment in frontoparietal regions has not been widely studied, despite general implication of these in top-down selection of information. To investigate this, we let participants perform an established intermodal selective attention task, where low-frequency auditory (1.6 Hz) and visual (1.8 Hz) stimuli were presented simultaneously. We instructed them to either attend to the auditory or to the visual stimuli and to detect targets while ignoring the other stimulus stream. As expected, the strongest phase adjustment was observed in primary sensory regions for auditory and for visual stimulation, independent of attentional focus. We found greater differences in phase locking between attended and ignored stimulation for the visual modality. Interestingly, auditory temporal regions show small but significant attention-dependent neural entrainment even for visual stimulation. Extending findings from invasive recordings in non-human primates, we demonstrate an effect of attentional focus on the phase of the entrained oscillations in auditory and visual cortex which may be driven by phase locked increases of induced power. While sensory areas adjusted the phase of the respective stimulation frequencies, attentional focus adjusted the peak frequencies in nonsensory areas. Spatially these areas show a striking overlap with core regions of the dorsal attention network and the frontoparietal network. This suggests that these areas prioritize the attended modality by optimally exploiting the temporal structure of stimulation. Overall, our study complements and extends previous work by showing a differential effect of attentional focus on entrained oscillations (or phase adjustment) in primary sensory areas and frontoparietal areas.

Dynamic pulvino-cortical interactions in the primate attention network.

While research in previous decades demonstrated a link between the pulvinar nucleus of the thalamus and visual selective attention, the pulvinar's specific functional role has remained elusive. However, methodological advances in electrophysiological recordings in non-human primates, including simultaneous recordings in multiple brain regions, have recently begun to reveal the pulvinar's functional contributions to selective attention. These new findings suggest that the pulvinar is critical for the efficient transmission of sensory information within and between cortical regions, both synchronizing cortical activity across brain regions and controlling cortical excitability. These new findings further suggest that the pulvinar's influence on cortical processing is embedded in a dynamic selection process that balances sensory and motor functions within the large-scale network that directs selective attention.

A surface metric and software toolbox for EEG electrode grids in the macaque

BackgroundThe past years have seen increased appreciation of electroencephalographic (EEG) recordings in non-human primates (NHP) as a tool for translational research. In humans, even large EEG electrode grids can easily and quickly be placed on standardized positions using commercially available EEG caps. In the NHP, the identification of standardized EEG electrode positions is more complicated and time-consuming. New methodHere we introduce a surface metric and software package (NHP1020) that automates the planning of large, approximately evenly spaced electrode grids for EEG recordings in the NHP. ResultsBased on one CT and one MRI image as well as two intracranial markers, the NHP1020 software defines electrode positions on the brain surface using a surface-based spherical metric similar to the one used by the international 10-20 system. Standardized electrode grids can be shared, imported or defined with few high-level commands. Existing methodsNHP EEG electrodes can be placed relative to extracranial markers and measurements or relative to underlying neural structures of interest. Both approaches are time-consuming and require manual intervention. Furthermore, the use of extracranial markers in this species may be more problematic than in humans, because cranial muscles and ridges are larger and keep maturing long into adulthood. ConclusionThe presented surface metric and the NHP1020 toolbox provide fast and automated identification of entire electrode grids in the non-human primate based on a two-dimensional metric on the brain surface.

Transcranial alternating current stimulation attenuates BOLD adaptation and increases functional connectivity.

Transcranial alternating current stimulation (tACS) is used as a noninvasive tool for cognitive enhancement and clinical applications. The physiological effects of tACS, however, are complex and poorly understood. Most studies of tACS focus on its ability to entrain brain oscillations, but our behavioral results in humans and extracellular recordings in nonhuman primates support the view that tACS at 10 Hz also affects brain function by reducing sensory adaptation. Our primary goal in the present study is to test this hypothesis using blood oxygen level-dependent (BOLD) imaging in human subjects. Using concurrent functional magnetic resonance imaging (fMRI) and tACS, and a motion adaptation paradigm developed to quantify BOLD adaptation, we show that tACS significantly attenuates adaptation in the human motion area (hMT+). In addition, an exploratory analysis shows that tACS increases functional connectivity of the stimulated hMT+ with the rest of the brain and the dorsal attention network in particular. Based on field estimates from individualized head models, we relate these changes to the strength of tACS-induced electric fields. Specifically, we report that functional connectivity (between hMT+ and any other region of interest) increases in proportion to the field strength in the region of interest. These findings add support for the claim that weak 10-Hz currents applied to the scalp modulate both local and global measures of brain activity.NEW & NOTEWORTHY Concurrent transcranial alternating current stimulation (tACS) and functional MRI show that tACS affects the human brain by attenuating adaptation and increasing functional connectivity in a dose-dependent manner. This work is important for our basic understanding of what tACS does, but also for therapeutic applications, which need insight into the full range of ways in which tACS affects the brain.

Neuronal signatures of a random-dot motion comparison task

The study of perceptual decision making has made significant progress owing to major contributions from two experimental paradigms: the sequential vibrotactile frequency comparison task for the somatosensory domain requiring working memory, and the random-dot motion task in the visual domain requiring evidence accumulation over time. On the one hand, electrophysiological recordings in nonhuman primates and humans have identified changes in firing rates and power modulations of beta band oscillations with the vibrotactile frequencies held in working memory, as well as with the mental operation required for decision making. On the other hand, firing rates and centro-parietal potentials were found to increase to a fixed level at the time of responding during the random-dot motion task, possibly reflecting an underlying evidence accumulation mechanism until a decision threshold is met. Here, to bridge these two paradigms, we presented two visual random-dot motion stimuli in a sequential comparison task while recording EEG from human volunteers. We identified a modulation of prefrontal beta band power that scaled with the level of dot motion coherence of the first stimulus during a short retention interval. Furthermore, beta power in premotor areas was modulated by participants’ choices approximately 700 ms before responses were given via button press. At the same time, dot motion patches of the second stimulus evoked a pattern of broadband centro-parietal signal build-up till responses were made, whose peak varied with trial difficulty. Hence, we show that known modulations of beta power during working memory and decision making extend from the vibrotactile to the visual domain and provide support for the notion of evidence accumulation as an unconfined decision-making mechanism generalizing over distinct decision types.

A wireless and artefact-free 128-channel neuromodulation device for closed-loop stimulation and recording in non-human primates.

Closed-loop neuromodulation systems aim to treat a variety of neurological conditions by delivering and adjusting therapeutic electrical stimulation in response to a patient's neural state, recorded in real time. Existing systems are limited by low channel counts, lack of algorithmic flexibility, and the distortion of recorded signals by large and persistent stimulation artefacts. Here, we describe an artefact-free wireless neuromodulation device that enables research applications requiring high-throughput data streaming, low-latency biosignal processing, and simultaneous sensing and stimulation. The device is a miniaturized neural interface capable of closed-loop recording and stimulation on 128 channels, with on-board processing to fully cancel stimulation artefacts. In addition, it can detect neural biomarkers and automatically adjust stimulation parameters in closed-loop mode. In a behaving non-human primate, the device enabled long-term recordings of local field potentials and the real-time cancellation of stimulation artefacts, as well as closed-loop stimulation to disrupt movement preparatory activity during a delayed-reach task. The neuromodulation device may help advance neuroscientific discovery and preclinical investigations of stimulation-based therapeutic interventions.

Right Hemisphere Lateralization in Neural Connectivity Within Fronto-Parietal Networks in Non-human Primates During a Visual Reaching Task.

A fronto-parietal network, comprised of the posterior parietal cortex (PPC) and the dorsal premotor cortex (PMd) has been proposed to be involved in planning and guiding movement. However, the issue of how the network is expressed across the bilateral cortical area according to the effector's side remains unclear. In this study, we tested these questions using electrocorticographic (ECoG) recordings in non-human primates and using a simple visual guided reaching task that induced a left or right hand response based on relevant cues provided for the task. The findings indicate that right hemisphere lateralized network patterns in which the right PMd was strongly coordinated with bilateral PPC immediately after presentation of the movement cue occurred, while the coherence with the left PMd was not enhanced. No difference was found in the coherence pattern between the effector's side (left hand or right hand), but the strength of coherence was different, in that animals showed a higher coherence in the right hand response compared to the left. Our data support that right lateralization in long-range phase synchrony in the 10–20 Hz low beta band is involved in motor preparation stage, irrespective of the upcoming effector's side.

Pharmacological inactivation does not support a unique causal role for intraparietal sulcus in the discrimination of visual number.

The “number sense” describes the intuitive ability to quantify without counting. Single neuron recordings in non-human primates and functional imaging in humans suggest the intraparietal sulcus is an important neuroanatomical locus of numerical estimation. Other lines of inquiry implicate the IPS in numerous other functions, including attention and decision making. Here we provide a direct test of whether IPS has functional specificity for numerosity judgments. We used muscimol to reversibly and independently inactivate the ventral and lateral intraparietal areas in two monkeys performing a numerical discrimination task and a color discrimination task, roughly equilibrated for difficulty. Inactivation of either area caused parallel impairments in both tasks and no evidence of a selective deficit in numerical processing. These findings do not support a causal role for the IPS in numerical discrimination, except insofar as it also has a role in the discrimination of color. We discuss our findings in light of several alternative hypotheses of IPS function, including a role in orienting responses, a general cognitive role in attention and decision making processes and a more specific role in ordinal comparison that encompasses both number and color judgments.

Characterization of neural entrainment to speech with and without slow spectral energy fluctuations in laminar recordings in monkey A1

Neural entrainment, the alignment between neural oscillations and rhythmic stimulation, is omnipresent in current theories of speech processing – nevertheless, the underlying neural mechanisms are still largely unknown. Here, we hypothesized that laminar recordings in non-human primates provide us with important insight into these mechanisms, in particular with respect to processing in cortical layers. We presented one monkey with human everyday speech sounds and recorded neural (as current-source density, CSD) oscillations in primary auditory cortex (A1). We observed that the high-excitability phase of neural oscillations was only aligned with those spectral components of speech the recording site was tuned to; the opposite, low-excitability phase was aligned with other spectral components. As low- and high-frequency components in speech alternate, this finding might reflect a particularly efficient way of stimulus processing that includes the preparation of the relevant neuronal populations to the upcoming input. Moreover, presenting speech/noise sounds without systematic fluctuations in amplitude and spectral content and their time-reversed versions, we found significant entrainment in all conditions and cortical layers. When compared with everyday speech, the entrainment in the speech/noise conditions was characterized by a change in the phase relation between neural signal and stimulus and the low-frequency neural phase was dominantly coupled to activity in a lower gamma-band. These results show that neural entrainment in response to speech without slow fluctuations in spectral energy includes a process with specific characteristics that is presumably preserved across species.