Identification of a stress-responsive subregion of the basolateral amygdala in male rats.

The basolateral amygdala (BLA) is reliably activated by psychological stress and hyperactive in conditions of pathological stress or trauma; however, subsets of BLA neurons are also readily activated by rewarding stimuli and can suppress fear and avoidance behaviours. The BLA is highly heterogeneous anatomically, exhibiting continuous molecular and connectivity gradients throughout the entire structure. A critical gap remains in understanding the anatomical specificity of amygdala subregions, circuits, and cell types explicitly activated by acute stress and how they are dynamically activated throughout stimulus exposure. Using a combination of topographical mapping for the activity-responsive protein FOS and fiber photometry to measure calcium transients in real-time, we sought to characterize the spatial and temporal patterns of BLA activation in response to a range of novel stressors (shock, swim, restraint, predator odour) and non-aversive, but novel stimuli (crackers, citral odour). We report four main findings: (1) the BLA exhibits clear spatial activation gradients in response to novel stimuli throughout the medial-lateral and dorsal-ventral axes, with aversive stimuli strongly biasing activation towards medial aspects of the BLA; (2) novel stimuli elicit distinct temporal activation patterns, with stressful stimuli exhibiting particularly enhanced or prolonged temporal activation patterns; (3) changes in BLA activity are associated with changes in behavioural state; and (4) norepinephrine enhances stress-induced activation of BLA neurons via the ß-noradrenergic receptor. Moving forward, it will be imperative to combine our understanding of activation gradients with molecular and circuit-specificity.

Covalently Attached Slippery Surface Coatings to Reduce Protein Adsorptions on Poly(dimethylsiloxane) Planar Surfaces and 3D Microfluidic Channels.

Silicone elastomers, such as poly(dimethylsiloxane) (PDMS), have a broad range of applications in basic biomedical research and clinical medicine, ranging from the preparation of microfluidic devices for organs-on-chips and ventriculoperitoneal shunts for the treatment of hydrocephalus to implantable neural probes for neuropharmacology. Despite the importance, the protein adsorptions on silicone elastomers in these application environments represent a significant challenge. Surface coatings with slippery lubricants, inspired by the Nepenthes pitcher plants, have recently received much attention for reducing protein adsorptions. Nevertheless, the depletion of the physically infused lubricants limits their broad applications. In this study, we report a covalently attached slippery surface coating to reduce protein adsorptions on PDMS surfaces. As demonstrations, we show that the adsorption of serum proteins, human fibrinogen and albumin, can be significantly reduced by the slippery surface coating in both planar PDMS surfaces and 3D microfluidic channels. The preparation of slippery surface coatings relies on the acid-catalyzed polycondensation reaction of dimethyldimethoxysilane, which utilizes a low-cost and scalable dip-coating method. Furthermore, cell metabolic activity and viability studies demonstrate the biocompatibility of the surface coating. These results suggest the potential applications of slippery surface coatings to reduce protein adsorptions for implantable medical devices, organs-on-chips, and many others.

Implantable Aptamer-Graphene Microtransistors for Real-Time Monitoring of Neurochemical Release in Vivo.

The real-time monitoring of neurochemical release in vivo plays a critical role in understanding the biochemical process of the complex nervous system. Current technologies for such applications, including microdialysis and fast-scan cyclic voltammetry, suffer from limited spatiotemporal resolution or poor selectivity. Here, we report a soft implantable aptamer-graphene microtransistor probe for real-time monitoring of neurochemical release. As a demonstration, we show the monitoring of dopamine with nearly cellular-scale spatial resolution, high selectivity (dopamine sensor >19-fold over norepinephrine), and picomolar sensitivity, simultaneously. Systematic benchtop evaluations, ex vivo experiments, and in vivo studies in mice models highlight the key features and demonstrate the capability of capturing the dopamine release dynamics evoked by pharmacological stimulation, suggesting the potential applications in basic neuroscience studies and studying neurological disease-related processes. The developed system can be easily adapted for monitoring other neurochemicals and drugs by simply replacing the aptamers functionalized on the graphene microtransistors.

Four methods of brain pattern analyses of fMRI signals associated with wrist extension versus wrist flexion studied for potential use in future motor learning BCI.

OBJECTIVE: In stroke survivors, a treatment-resistant problem is inability to volitionally differentiate upper limb wrist extension versus flexion. When one intends to extend the wrist, the opposite occurs, wrist flexion, rendering the limb non-functional. Conventional therapeutic approaches have had limited success in achieving functional recovery of patients with chronic and severe upper extremity impairments. Functional magnetic resonance imaging (fMRI) neurofeedback is an emerging strategy that has shown potential for stroke rehabilitation. There is a lack of information regarding unique blood-oxygenation-level dependent (BOLD) cortical activations uniquely controlling execution of wrist extension versus uniquely controlling wrist flexion. Therefore, a first step in providing accurate neural feedback and training to the stroke survivor is to determine the feasibility of classifying (or differentiating) brain activity uniquely associated with wrist extension from that of wrist flexion, first in healthy adults. APPROACH: We studied brain signal of 10 healthy adults, who performed wrist extension and wrist flexion during fMRI data acquisition. We selected four types of analyses to study the feasibility of differentiating brain signal driving wrist extension versus wrist flexion, as follows: 1) general linear model (GLM) analysis; 2) support vector machine (SVM) classification; 3) 'Winner Take All'; and 4) Relative Dominance. RESULTS: With these four methods and our data, we found that few voxels were uniquely active during either wrist extension or wrist flexion. SVM resulted in only minimal classification accuracies. There was no significant difference in activation magnitude between wrist extension versus flexion; however, clusters of voxels showed extension signal > flexion signal and other clusters vice versa. Spatial patterns of activation differed among subjects. SIGNIFICANCE: We encountered a number of obstacles to obtaining clear group results in healthy adults. These obstacles included the following: high variability across healthy adults in all measures studied; close proximity of uniquely active voxels to voxels that were common to both the extension and flexion movements; in general, higher magnitude of signal for the voxels common to both movements versus the magnitude of any given uniquely active voxel for one type of movement. Our results indicate that greater precision in imaging will be required to develop a truly effective method for differentiating wrist extension versus wrist flexion from fMRI data.

Combined real-time fMRI and real time fNIRS brain computer interface (BCI): Training of volitional wrist extension after stroke, a case series pilot study.

OBJECTIVE: Pilot testing of real time functional magnetic resonance imaging (rt-fMRI) and real time functional near infrared spectroscopy (rt-fNIRS) as brain computer interface (BCI) neural feedback systems combined with motor learning for motor recovery in chronic severely impaired stroke survivors. APPROACH: We enrolled a four-case series and administered three sequential rt-fMRI and ten rt-fNIRS neural feedback sessions interleaved with motor learning sessions. Measures were: Arm Motor Assessment Tool, functional domain (AMAT-F; 13 complex functional tasks), Fugl-Meyer arm coordination scale (FM); active wrist extension range of motion (ROM); volume of activation (fMRI); and fNIRS HbO concentration. Performance during neural feedback was assessed, in part, using percent successful brain modulations during rt-fNIRS. MAIN RESULTS: Pre-/post-treatment mean clinically significant improvement in AMAT-F (.49 ± 0.22) and FM (10.0 ± 3.3); active wrist ROM improvement ranged from 20° to 50°. Baseline to follow-up change in brain signal was as follows: fMRI volume of activation was reduced in almost all ROIs for three subjects, and for one subject there was an increase or no change; fNIRS HbO was within normal range, except for one subject who increased beyond normal at post-treatment. During rt-fNIRS neural feedback training, there was successful brain signal modulation (42%-78%). SIGNIFICANCE: Severely impaired stroke survivors successfully engaged in spatially focused BCI systems, rt-fMRI and rt-fNIRS, to clinically significantly improve motor function. At the least, equivalency in motor recovery was demonstrated with prior long-duration motor learning studies (without neural feedback), indicating that no loss of motor improvement resulted from substituting neural feedback sessions for motor learning sessions. Given that the current neural feedback protocol did not prevent the motor improvements observed in other long duration studies, even in the presence of fewer sessions of motor learning in the current work, the results support further study of neural feedback and its potential for recovery of motor function in stroke survivors. In future work, expanding the sophistication of either or both rt-fMRI and rt-fNIRS could hold the potential for further reducing the number of hours of training needed and/or the degree of recovery. ClinicalTrials.gov ID: NCT02856035.

Development of a combined, sequential real-time fMRI and fNIRS neurofeedback system to enhance motor learning after stroke.

BACKGROUND: After stroke, wrist extension dyscoordination precludes functional arm/hand. We developed a more spatially precise brain signal for use in brain computer interface (BCI's) for stroke survivors. NEW METHOD: Combination BCI protocol of real-time functional magnetic resonance imaging (rt-fMRI) sequentially followed by functional near infrared spectroscopy (rt-fNIRS) neurofeedback, interleaved with motor learning sessions without neural feedback. Custom Matlab and Python code was developed to provide rt-fNIRS-based feedback to the chronic stroke survivor, system user. RESULTS: The user achieved a maximum of 71 % brain signal accuracy during rt-fNIRS neural training; progressive focus of brain activation across rt-fMRI neural training; increasing trend of brain signal amplitude during wrist extension across rt-fNIRS training; and clinically significant recovery of arm coordination and active wrist extension. COMPARISON WITH EXISTING METHODS: Neurorehabilitation, peripherally directed, shows limited efficacy, as do EEG-based BCIs, for motor recovery of moderate/severely impaired stroke survivors. EEG-based BCIs are based on electrophysiological signal; whereas, rt-fMRI and rt-fNIRS are based on neurovascular signal. CONCLUSION: The system functioned well during user testing. Methods are detailed for others' use. The system user successfully engaged rt-fMRI and rt-fNIRS neurofeedback systems, modulated brain signal during rt-fMRI and rt-fNIRS training, according to volume of brain activation and intensity of signal, respectively, and clinically significantly improved limb coordination and active wrist extension. fNIRS use in this case demonstrates a feasible/practical BCI system for further study with regard to use in chronic stroke rehab, and fMRI worked in concept, but cost and some patient-use issues make it less feasible for clinical practice.