Publications using TDT

Neurological disorders affect millions of Americans and this number is expected to rise with the aging population. Development of drugs to treat these disorders may be facilitated by improved in vitro models that faithfully reproduce salient features of the relevant brain regions. Current 3D culture methods face challenges with reliably reproducing microarchitectural features of brain morphology such as cortical or hippocampal layers. In this work, polydimethylsiloxane (PDMS) mini-wells were used to create low aspect ratio, adherent 3D constructs where neurons self-assemble into layers. Layer self-assembly was determined to depend on the size of the PDMS mini-well. Layer formation occurred in cultures composed of primary rat cortical neurons or human induced pluripotent stem cell—derived neurons and astrocytes and was robust and reproducible. Layered 3D constructs were found to have spontaneous neural activity characterized by long bursts similar to activity in the developing cortex. The responses of layered 3D cultures to drugs were more similar to in vivo data than those of 2D cultures. 3D constructs created with this method may be thus suitable as in vitro models for drug discovery for neurological disorders.

Bilateral cochlear implantation has provided access to some of the benefits of binaural hearing enjoyed by normal-hearing (NH) listeners. However, a gap in performance still exists between the two populations. Single-channel stimulation studies have shown that interaural place-of-stimulation mismatch (IPM) due to differences in implantation depth leads to decreased binaural fusion and lateralization of interaural time and level differences (ITDs and ILDs, respectively). While single-channel studies are informative, multi-channel stimulation is needed for good speech understanding with cochlear implants (CIs). Some multi-channel studies have shown that channel interaction due to current spread can affect ITD sensitivity. In this work, we studied the effect of IPM and channel spacing, along with their potential interaction, on binaural fusion and ITD/ILD lateralization. Experiments were conducted in adult NH listeners and CI listeners with a history of acoustic hearing. Results showed that IPM reduced the range of lateralization for ITDs but not ILDs. CI listeners were more likely to report a fused percept in the presence of IPM with multi-channel stimulation than NH listeners. However, no effect of channel spacing was found. These results suggest that IPM should be accounted for in clinical mapping practices in order to maximize bilateral CI benefits.

Separating sound sources in acoustic environments relies on making ongoing, highly accurate spectro-temporal comparisons. However, listeners with hearing impairment may have varying quality of temporal encoding within or across ears, which may limit the listeners' ability to make spectro-temporal comparisons between places-of-stimulation. In this study in normal hearing listeners, depth of amplitude modulation (AM) for sinusoidally amplitude modulated (SAM) tones was manipulated in an effort to reduce the coding of periodicity in the auditory nerve. The ability to judge differences in AM rates was studied for stimuli presented to different cochlear places-of-stimulation, within- or across-ears. It was hypothesized that if temporal encoding was poorer for one tone in a pair, then sensitivity to differences in AM rate of the pair would decrease. Results indicated that when the depth of AM was reduced from 50% to 20% for one SAM tone in a pair, sensitivity to differences in AM rate decreased. Sensitivity was greatest for AM rates near 90 Hz and depended upon the places-of-stimulation being compared. These results suggest that degraded temporal representations in the auditory nerve for one place-of-stimulation could lead to deficits comparing that temporal information with other places-of-stimulation.

Background: Rhesus macaques are the most popular model species for studying the neural basis of visual face processing and social interaction using intracranial methods. However, the challenge of creating realistic, dynamic, and parametric macaque face stimuli has limited the experimental control and ethological validity of existing approaches. New method: We performed statistical analyses of in vivo computed tomography data to generate an anatomically accurate, three-dimensional representation of Rhesus macaque cranio-facial morphology. The surface structures were further edited, rigged and textured by a professional digital artist with careful reference to photographs of macaque facial expression, colouration and pelage. Results: The model offers precise, continuous, parametric control of craniofacial shape, emotional expression, head orientation, eye gaze direction, and many other parameters that can be adjusted to render either static or dynamic high-resolution faces. Example single-unit responses to such stimuli in macaque inferotemporal cortex demonstrate the value of parametric control over facial appearance and behaviours. Comparison with existing method(s): The generation of such a high-dimensionality and systematically controlled stimulus set of conspecific faces, with accurate craniofacial modelling and professional finalization of facial details, is currently not achievable using existing methods. Conclusions: The results herald a new set of possibilities in adaptive sampling of a high-dimensional and socially meaningful feature space, thus opening the door to systematic testing of hypotheses about the abundant neural specialization for faces found in the primate.

Previous studies have demonstrated that pairing vagus nerve stimulation (VNS) with sounds can enhance the primary auditory cortex (A1) response to the paired sound. The neural response to sounds following VNS-sound pairing in other subcortical and cortical auditory fields has not been documented. We predicted that VNS-tone pairing would increase neural responses to the paired tone frequency across the auditory pathway. In this study, we paired VNS with the presentation of a 9-kHz tone 300 times a day for 20 days. We recorded neural responses to tones from 2,950 sites in the inferior colliculus (IC), A1, anterior auditory field (AAF), and posterior auditory field (PAF) 24 h after the last pairing session in anesthetized rats. We found that VNS-tone pairing increased the percentage of IC, A1, AAF, and PAF that responds to the paired tone frequency. Across all tested auditory fields, the response strength to tones was strengthened in VNS-tone paired rats compared with control rats. VNS-tone pairing reduced spontaneous activity, frequency selectivity, and response threshold across the auditory pathway. This is the first study to document both cortical and subcortical plasticity following VNS-sound pairing. Our findings suggest that VNS paired with sound presentation is an effective method to enhance auditory processing. NEW & NOTEWORTHY Previous studies have reported primary auditory cortex plasticity following vagus nerve stimulation (VNS) paired with a sound. This study extends previous findings by documenting that fields across the auditory pathway are altered by VNS-tone pairing. VNS-tone pairing increases the percentage of each field that responds to the paired tone frequency. This is the first study to document both cortical and subcortical plasticity following VNS-sound pairing.

To make adaptive decisions, organisms must appropriately filter sensory inputs, augmenting relevant signals and suppressing noise. The prefrontal cortex (PFC) partly implements this process by regulating thalamic activity through modality-specific thalamic reticular nucleus (TRN) subnetworks. However, because the PFC does not directly project to sensory TRN subnetworks, the circuitry underlying this process had been unknown. Here, using anatomical tracing, functional manipulations, and optical identification of PFC projection neurons, we find that the PFC regulates sensory thalamic activity through a basal ganglia (BG) pathway. Engagement of this PFC-BG-thalamus pathway enables selection between vision and audition by primarily suppressing the distracting modality. This pathway also enhances sensory discrimination and is used for goal-directed background noise suppression. Overall, our results identify a new pathway for attentional filtering and reveal its multiple roles in sensory processing on the basis of internal goals.

The explore-exploit dilemma refers to the challenge of deciding when to forego immediate rewards and explore new opportunities that could lead to greater rewards in the future. While motivational neural circuits facilitate learning based on past choices and outcomes, it is unclear whether they also support computations relevant for deciding when to explore. We recorded neural activity in the amygdala and ventral striatum of rhesus macaques as they solved a task that required them to balance novelty-driven exploration with exploitation of what they had already learned. Using a partially observable Markov decision process (POMDP) model to quantify explore-exploit trade-offs, we identified that the ventral striatum and amygdala differ in how they represent the immediate value of exploitative choices and the future value of exploratory choices. These findings show that subcortical motivational circuits are important in guiding explore-exploit decisions.

The CHRNA7 gene that encodes the α 7 -subunit of the nicotinic acetylcholine receptor (α 7 -nAChR) has been associated with some autism spectrum disorders and other neurodevelopmental conditions characterized, in part, by auditory and language impairment. These conditions may include auditory processing disorders that represent impaired timing of neural activity, often accompanied by problems understanding speech. Here, we measure timing properties of sound-evoked activity via the auditory brainstem response (ABR) of α 7 -nAChR knockout mice of both sexes and wild-type colony controls. We find a significant timing delay in evoked ABR signals that represents midbrain activity in knockouts. We also examine spike-timing properties of neurons in the inferior colliculus, a midbrain nucleus that exhibits high levels of α 7 -nAChR during development. We find delays of evoked responses along with degraded spiking precision in knockout animals. We find similar timing deficits in responses of neurons in the superior paraolivary nucleus and ventral nucleus of the lateral lemniscus, which are brainstem nuclei thought to shape temporal precision in the midbrain. In addition, we find that other measures of temporal acuity including forward masking and gap detection are impaired for knockout animals. We conclude that altered temporal processing at the level of the brainstem in α 7 -nAChR-deficient mice may contribute to degraded spike timing in the midbrain, which may underlie the observed timing delay in the ABR signals. Our findings are consistent with a role for the α 7 -nAChR in types of neurodevelopmental and auditory processing disorders and we identify potential neural targets for intervention. NEW & NOTEWORTHY Disrupted signaling via the α 7 -nicotinic acetylcholine receptor (α 7 -nAChR) is associated with neurodevelopmental disorders that include impaired auditory processing. The underlying causes of dysfunction are not known but a common feature is abnormal timing of neural activity. We examined temporal processing of α 7 -nAChR knockout mice and wild-type controls. We found degraded spike timing of neurons in knockout animals, which manifests at the level of the auditory brainstem and midbrain.

Bone remodeling of the auditory ossicles and the otic capsule is highly restricted and tightly controlled by the osteoprotegerin (OPG)/receptor activator of nuclear factor kappa-Β ligand (RANKL)/receptor activator of nuclear factor kappa-Β (RANK) system. In these bony structures, a pathological decrease in OPG expression stimulates osteoclast differentiation and excessive resorption followed by accrual of sclerotic bone, ultimately resulting in the development of otosclerosis, a leading cause of deafness in adults. Understanding the signaling pathways involved in maintaining OPG expression in the ear would shed light on the pathophysiology of otosclerosis and other ear bone-related diseases. We and others previously demonstrated that Ca2+ signaling through the L-type CaV1.2 Ca2+ channel positively regulates OPG expression and secretion in long bone osteoblasts and their precursor cells in vitro and in vivo. Whether CaV1.2 regulates OPG expression in ear bones has not been investigated. We drove expression of a gain-of-function CaV1.2 mutant channel (CaV1.2TS) using Col2a1-Cre, which we found to target osteochondral/osteoblast progenitors in the auditory ossicles and the otic capsule. Col2a1-Cre;CaV1.2TS mice displayed osteopetrosis of these bones shown by μCT 3D reconstruction, histological analysis, and lack of bone sculpting, findings similar to phenotypes seen in mice with an osteoclast defect. Consistent with those observations, we found that Col2a1-Cre;CaV1.2TS mutant mice showed reduced osteoclasts in the otic capsule, upregulated mRNA expression of Opg and Opg/Rankl ratio, and increased mRNA expression of osteoblast differentiation marker genes in the otic capsule, suggesting both an anti-catabolic and anabolic effect of CaV1.2TS mutant channel contributed to the observed morphological changes of the ear bones. Further, we found that Col2a1-Cre;CaV1.2TS mice experienced hearing loss and displayed defects of body balance in behavior tests, confirming that the CaV1.2-dependent Ca2+ influx affects bone structure in the ear and consequent hearing and vestibular functions. Together, these data support our hypothesis that Ca2+ influx through CaV1.2TS promotes OPG expression from osteoblasts, thereby affecting bone modeling/remodeling in the auditory ossicles and the otic capsule. These data provide insight into potential pathological mechanisms underlying perturbed OPG expression and otosclerosis.

Functional magnetic resonance imaging (fMRI) is commonly thought to be too slow to capture any neural dynamics faster than 0.1 Hz. However, recent findings demonstrate the feasibility of detecting fMRI activity at higher frequencies beyond 0.2 Hz. The origin, reliability, and generalizability of fast fMRI responses are still under debate and await confirmation through animal experiments with fMRI and invasive electrophysiology. Here, we acquired single-echo and multi-echo fMRI, as well as local field potentials, from anesthetized rat brains given gastric electrical stimulation modulated at 0.2, 0.4 and 0.8 Hz. Such gastric stimuli could drive widespread fMRI responses at corresponding frequencies from the somatosensory and cingulate cortices. Such fast fMRI responses were linearly dependent on echo times and thus indicative of blood oxygenation level dependent nature (BOLD). Local field potentials recorded during the same gastric stimuli revealed transient and phase-locked broadband neural responses, preceding the fMRI responses by as short as 0.5 s. Taken together, these results suggest that gastric stimulation can drive widespread and rapid fMRI responses of BOLD and neural origin, lending support to the feasibility of using fMRI to detect rapid changes in neural activity up to 0.8 Hz under visceral stimulation.

A current challenge of neurotechnologies is the development of speech brain-computer interfaces to restore communication in people unable to speak. To achieve a proof of concept of such system, neural activity of patients implanted for clinical reasons can be recorded while they speak. Using such simultaneously recorded audio and neural data, decoders can be built to predict speech features using features extracted from brain signals. A typical neural feature is the spectral power of field potentials in the high-gamma frequency band (between 70 and 200 Hz), a range that happens to overlap the fundamental frequency of speech. Here, we analyzed human electrocorticographic (ECoG) and intracortical recordings during speech production and perception as well as rat microelectrocorticographic (µ-ECoG) recordings during sound perception. We observed that electrophysiological signals, recorded with different recording setups, often contain spectrotemporal features highly correlated with those of the sound, especially within the high-gamma band. The characteristics of these correlated spectrotemporal features support a contamination of electrophysiological recordings by sound. In a recording showing high contamination, using neural features within the high-gamma frequency band dramatically increased the performance of linear decoding of acoustic speech features, while such improvement was very limited for another recording showing weak contamination. Further analysis and in vitro replication suggest that the contamination is caused by a mechanical action of the sound waves onto the cables and connectors along the recording chain, transforming sound vibrations into an undesired electrical noise that contaminates the biopotential measurements. This study does not question the existence of relevant physiological neural information underlying speech production or sound perception in the high-gamma frequency band, but alerts on the fact that care should be taken to evaluate and eliminate any possible acoustic contamination of neural signals in order to investigate the cortical dynamics of these processes.

Cortical encoding of auditory space relies on two major peripheral cues, interaural time difference (ITD) and interaural level difference (ILD) of the sounds arriving at a listener’s ears. In much of the precortical auditory pathway, ITD and ILD cues are processed independently, and it is assumed that cue integration is a higher order process. However, there remains debate on how ITDs and ILDs are encoded in the cortex and whether they share a common mechanism. The present study used electroencephalography (EEG) to measure evoked cortical potentials from narrowband noise stimuli with imposed binaural cue changes. Previous studies have similarly tested ITD shifts to demonstrate that neural populations broadly favor one spatial hemifield over the other, which is consistent with an opponent-channel model that computes the relative activity between broadly tuned neural populations. However, it is still a matter of debate whether the same coding scheme applies to ILDs and, if so, whether processing the two binaural cues is distributed across similar regions of the cortex. The results indicate that ITD and ILD cues have similar neural signatures with respect to the monotonic responses to shift magnitude; however, the direction of the shift did not elicit responses equally across cues. Specifically, ITD shifts evoked greater responses for outward than inward shifts, independently of the spatial hemifield of the shift, whereas ILD-shift responses were dependent on the hemifield in which the shift occurred. Active cortical structures showed only minor overlap between responses to cues, suggesting the two are not represented by the same pathway. NEW & NOTEWORTHY Interaural time differences (ITDs) and interaural level differences (ILDs) are critical to locating auditory sources in the horizontal plane. The higher order perceptual feature of auditory space is thought to be encoded together by these binaural differences, yet evidence of their integration in cortex remains elusive. Although present results show some common effects between the two cues, key differences were observed that are not consistent with an ITD-like opponent-channel process for ILD encoding.

Objective: The objective of the study was to quantify effective connectivity from the piriform cortex to mediodorsal thalamus, in Genetic Absence Epilepsy Rats from Strasbourg (GAERS). Methods: Local field potentials (LFPs) were recorded using microelectrode arrays implanted in the mediodorsal thalamus and piriform cortex, in three urethane anesthetized GAERS and three control rats. Screw electrodes were placed in the primary motor cortex to identify epileptiform discharges. We used transfer entropy to measure effective connectivity from piriform cortex to mediodorsal thalamus prior to and during generalized epileptiform discharges. Results: We observed increased theta band effective connectivity from piriform cortex to mediodorsal thalamus, prior to and during epileptiform discharges in GAERS compared with controls. Increased effective connectivity was also observed in beta and gamma bands from the piriform cortex to mediodorsal thalamus, but only during epileptiform discharges. Conclusions: This preliminary study suggests that increased effective theta connectivity from the piriform cortex to the mediodorsal thalamus may be a feature of the 'epileptic network' associated with genetic absence epilepsy. Our findings indicate an underlying predisposition of this direct pathway to propagate epileptiform discharges in genetic absence epilepsy.

Despite a wealth of clinical and preclinical data implicating the serotonin (5-HT) system in fear-related affective disorders, a precise definition of this neuromodulator's role in fear remains elusive. Using convergent anatomical and functional approaches, we interrogate the contribution to fear of basal amygdala (BA) 5-HT inputs from the dorsal raphe nucleus (DRN). We show the DRN→BA 5-HT pathway is engaged during fear memory formation and retrieval, and activity of these projections facilitates fear and impairs extinction. The DRN→BA 5-HT pathway amplifies fear-associated BA neuronal firing and theta power and phase-locking. Although fear recruits 5-HT and VGluT3 co-expressing DRN neurons, the fear-potentiating influence of the DRN→BA 5-HT pathway requires signaling at BA 5-HT1A/2A receptors. Input-output mapping illustrates how the DRN→BA 5-HT pathway is anatomically distinct and connected with other brain regions that mediate fear. These findings reveal how a discrete 5-HT circuit orchestrates a broader neural network to calibrate aversive memory.

An experiment was performed with 10 young normal-hearing listeners that attempted to determine if envelope modulations affected binaural processing in bandlimited pulse trains. Listeners detected an interaurally out-of-phase carrier pulse train in the presence of different amplitude modulations. The peaks of the pulses were constant (called "flat" or F), followed envelope modulations from an interaurally correlated 50-Hz bandwidth noise (called CM), or followed modulations from an interaurally uncorrelated noise (called UM). The pulse rate was varied from 50 to 500 pulses per second (pps) and the center frequency (CF) was 4 or 8 kHz. It was hypothesized that CM would cause no change or an increase in performance compared to F; UM would cause a decrease because of the blurring of the binaural detection cue. There was a small but significant decrease from F to CM (inconsistent with the hypothesis) and a further decrease from CM to UM (consistent with the hypothesis). Critically, there was a significant envelope by rate interaction caused by a decrease from F to CM for the 200-300 pps rates. The data can be explained by a subject-based factor, where some listeners experienced interaural envelope decorrelation when the sound was encoded by the auditory system that reduced performance when the modulations were present. Since the decrease in performance between F and CM conditions was small, it seems that most young normal-hearing listeners have very similar encoding of modulated stimuli across the ears. This type of task, when further optimized, may be able to assess if hearing-impaired populations experience interaural decorrelation from encoding modulated stimuli and therefore could help better understand the limited spatial hearing in populations like cochlear-implant users.

Although the effects of intense noise exposure on the peripheral and central auditory pathway have been well characterized, its effects on non-classical auditory structures in the brain, such as the hippocampus, are less well understood. Previously, we demonstrated that noise-induced hearing loss causes a significant long-term reduction in hippocampal neurogenesis and cell proliferation. Given the known suppressive effects of stress hormones on neurogenesis, the goal of the present study was to determine if activation of the stress response is an underlying mechanism for the long-term reduction in hippocampal neurogenesis observed following noise trauma. To accomplish this, we monitored basal and reactive blood plasma levels of the stress hormone corticosterone in rats for ten weeks following acoustic trauma, and quantified changes in hippocampal glucocorticoid and mineralocorticoid receptors. Our results indicate that long-term auditory deprivation does not cause a persistent increase in basal or reactive stress hormone levels in the weeks following noise exposure. Instead, we observed a greater decline in reactive corticosterone release in noise-exposed rats between the first and tenth week of sampling compared to control rats. We also observed a significant increase in hippocampal glucocorticoid receptor expression which may cause greater hippocampal sensitivity to circulating glucocorticoid levels and result in glucocorticoid-induced suppression of neurogenesis, as well as increased feedback inhibition on the HPA axis. No change in mineralocorticoid receptor expression was observed between control and noise exposed rats. These results highlight the adverse effect of intense noise exposure and auditory deprivation on the hippocampus.

Significance The firing dynamics of neurons in primary visual cortex (V1) are poorly understood. Indeed, V1 neurons of freely behaving rats fire at the same mean rate in light and darkness. It is unclear how this stability is maintained and whether it is important for sensory processing. We find that transitions between light and darkness happening at expected times have only modest effects on V1 activity. In contrast, both unexpected transitions and light reexposure after extended darkness robustly increase V1 firing. Finally, pairwise correlations in neuronal spiking are significantly higher during the light when natural vision is occurring. These data show that V1 firing rates are actively stabilized while simultaneously allowing for input-dependent changes in correlations between neurons.

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.

The rostromedioventral striatum is critical for behavior dependent on evaluating rewards. We asked what contribution tonically active neurons (TANs), the putative striatal cholinergic interneurons, make in coding reward value in this part of the striatum. Two female monkeys were given the option to accept or reject an offered reward in each trial, the value of which was signaled by a visual cue. Forty-five percent of the TANs use temporally modulated activity to encode information about discounted value. These responses were significantly better represented using principal component analysis than by just counting spikes. The temporal coding is straightforward: the spikes are distributed according to a sinusoidal envelope of activity that changes gain, ranging from positive to negative according to discounted value. Our results show that the information about the relative value of an offered reward is temporally encoded in neural spike trains of TANs. This temporal coding may allow well tuned, coordinated behavior to emerge.SIGNIFICANCE STATEMENT Ever since the discovery that neurons use trains of pulses to transmit information, it seemed self-evident that information would be encoded into the pattern of the spikes. However, there is not much evidence that spike patterns encode cognitive information. We find that a set of interneurons, the tonically active neurons (TANs) in monkeys' striatum, use temporal patterns of response to encode information about the discounted value of offered rewards. The code seems straightforward: a sinusoidal envelope that changes gain according to the discounted value of the offer, describes the rate of spiking across time. This temporal modulation may provide a means to synchronize these interneurons and the activity of other neural elements including principal output neurons.

Natural communication often occurs in dialogue, differentially engaging auditory and sensorimotor brain regions during listening and speaking. However, previous attempts to decode speech directly from the human brain typically consider listening or speaking tasks in isolation. Here, human participants listened to questions and responded aloud with answers while we used high-density electrocorticography (ECoG) recordings to detect when they heard or said an utterance and to then decode the utterance’s identity. Because certain answers were only plausible responses to certain questions, we could dynamically update the prior probabilities of each answer using the decoded question likelihoods as context. We decode produced and perceived utterances with accuracy rates as high as 61% and 76%, respectively (chance is 7% and 20%). Contextual integration of decoded question likelihoods significantly improves answer decoding. These results demonstrate real-time decoding of speech in an interactive, conversational setting, which has important implications for patients who are unable to communicate.