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Many experiments record sequential trajectories where each trajectory consists of oscillations and fluctuations around zero. Such trajectories can be viewed as zero-mean functional data. When there are structural breaks (on the sequence of trajectories) in higher order moments, it is not always easy to spot these by mere visual inspection. Motivated by this challenging problem in brain signal analysis, we propose a detection and testing procedure to find the change point in functional covariance. The detection procedure is based on the cumulative sum statistics (CUSUM). The classical testing procedure for functional data depends on a null distribution which depends on infinitely many unknown parameters, though in practice only a finite number of these can be included for the hypothesis test of the existence of change point. This paper provides some theoretical insights on the influence of the number of parameters. Meanwhile, the asymptotic properties of the estimated change point are developed. The effectiveness of the proposed method is numerically validated in simulation studies and an application to investigate changes in rat brain signals following an experimentally-induced stroke.

We have previously demonstrated protection from impending cortical stroke is achievable by sensory stimulation of the ischemic area in an adult rat model of permanent middle cerebral artery occlusion (pMCAo). We have further demonstrated that a major underpinning mechanism that is necessary for such protection is the system of collaterals among cerebral arteries that results in reperfusion of the MCA ischemic territory. However, since such collateral flow is weak, it may be necessary but not sufficient for protection and therefore we were seeking other complementary mechanisms that contribute to sensory-based protection. We hypothesized that astrocytes-to-neuron shuttle (ANLS) is another potential underpinning mechanism that could complement collateral flow in the protection process. Supporting our hypothesis, using functional imaging, pharmacological treatments, and postmortem histology, we show that ANLS has a pivotal role in sensory-based protection of cortex and therefor serves as the other supporting mechanism underpinning the protection process.

A new classification method for functional data is proposed in this paper. This work is motivated by the need to identify features that discriminate between neurological conditions on which local field potentials (LFPs) were recorded. Regardless of the condition, these local field potentials have zero mean and thus the first moments of these random processes do not have discriminating power. We propose the variation pattern classification (VPC) method {which employs the (auto-)covariance operators as the discriminating features} and uses the Hilbert-Schmidt norm to measure the discrepancy between the (auto-)covariance operators of different groups. The proposed VPC method is demonstrated to be sensitive to the discrepancy, {potentially leading to a higher rate of classification}. One important innovation lies in the dimension reduction where the VPC method data-adaptively determines the basis functions (discriminative feature functions) that account for the major discrepancy. In addition, the selected discriminative feature functions provide insights on the discrepancy between different groups because they reveal the features of variation pattern that differentiate groups. Consistency properties are established and, furthermore, simulation studies and the analysis of rat brain LFP trajectories empirically demonstrate the advantages and effectiveness of the proposed method.

Anesthetics, commonly used in preclinical and fundamental scientific research, have a depressive influence on the metabolic, neuronal, and vascular functions of the brain and can adversely influence neurophysiological results. The use of awake animals for research studies is advantageous but poses the major challenge of keeping the animals calm and stationary to minimize motion artifacts throughout data acquisition. Awake imaging in smaller-sized rodents (e.g., mice) is very common but remains scant in rats as rats are bigger, stronger, and have a greater tendency to oppose movement restraints and head fixation over the long durations required for imaging. A new model of neuroimaging of awake, head-fixed rats using customized hand-sewn slings, 3D-printed head implants, head caps, and a headframe is described. The results acquired following a single trial of single-whisker stimulation suggest an increase in the intensity of the evoked functional response. The acquisition of the evoked functional response from awake, head-fixed rats is faster than that from anesthetized rats, reliable, reproducible, and can be used for repeated longitudinal studies.

We propose a novel regularized mixture model for clustering matrix-valued data. The proposed method assumes a separable covariance structure for each cluster and imposes a sparsity structure (eg, low rankness, spatial sparsity) for the mean signal of each cluster. We formulate the problem as a finite mixture model of matrix-normal distributions with regularization terms, and then develop an expectation maximization type of algorithm for efficient computation. In theory, we show that the proposed estimators are strongly consistent for various choices of penalty functions. Simulation and two applications on brain signal studies confirm the excellent performance of the proposed method including a better prediction accuracy than the competitors and the scientific interpretability of the solution.

The goal of this paper is to develop a measure for characterizing complex dependence between time series that cannot be captured by traditional measures such as correlation and coherence. Our approach is to use copula models of functionals of the Fourier coefficients which is a generalization of coherence. Here, we use standard parametric copula models with a single parameter from both elliptical and Archimedean families. Our approach is to analyze changes in activity in local field potentials in the rat cortex prior to and immediately following the onset of stroke. We present the necessary theoretical background, the multivariate models and an illustration of our methodology on these local field potential data. Simulations with nonlinear dependent data reveal that there is information that is missed by not taking into account dependence on specific frequencies. Moreover, these simulations demonstrate how our proposed method captures more complex nonlinear dependence between time series. Finally, we illustrate our copula-based approach in the analysis of local field potentials of rats.

In a rat model of ischemic stroke by permanent occlusion of the medial cerebral artery (pMCAo), we have demonstrated using continuous recordings by microelectrode array at the depth of the ischemic territory that there is an immediate wide-spread increase in spontaneous local field potential synchrony following pMCAo that was correlated with ischemic stroke damage, but such increase was not seen in control sham-surgery rats. We further found that the underpinning source of the synchrony increase is intermittent bursts of low multi-frequency oscillations. Here we show that such increase in spontaneous LFP synchrony after pMCAo can be reduced to pre-pMCAo baseline level by delivering early (immediately after pMCAo) protective sensory stimulation that reduced the underpinning bursts. However, the delivery of a late (3 h after pMCAo) destructive sensory stimulation had no influence on the elevated LFP synchrony and its underpinning bursts. Histology confirmed both protection for the early stimulation group and an infarct for the late stimulation group. These findings highlight the unexpected importance of spontaneous LFP and its synchrony as a predictive correlate of cerebral protection or stroke infarct during the hyperacute state following pMCAo and the potential clinical relevance of stimulation to reduce EEG synchrony in acute stroke.

Real-time tracking of neurotransmitter levels in vivo has been technically challenging due to the low spatiotemporal resolution of current methods. Since the imbalance of cortical excitation/inhibition (E:I) ratios are associated with a variety of neurological disorders, accurate monitoring of excitatory and inhibitory neurotransmitter levels is crucial for investigating the underlying neural mechanisms of these conditions. Specifically, levels of the excitatory neurotransmitter L-glutamate, and the inhibitory neurotransmitter GABA, are assumed to play critical roles in the E:I balance. Therefore, in this work, a flexible electrochemical microsensor is developed for real-time simultaneous detection of L-glutamate and GABA. The flexible polyimide substrate was used for easier handling during implantation and measurement, along with less brain damage. Further, by electrochemically depositing Pt-black nanostructures on the sensor's surface, the active surface area was enhanced for higher sensitivity. This dual neurotransmitter sensor probe was validated under various settings for its performance, including in vitro, ex vivo tests with glutamatergic neuronal cells and in vivo test with anesthetized rats. Additionally, the sensor's performance has been further investigated in terms of longevity and biocompatibility. Overall, our dual L-glutamate:GABA sensor microprobe has its unique features to enable accurate, real-time, and long-term monitoring of the E:I balance in vivo. Thus, this new tool should aid investigations of neural mechanisms of normal brain function and various neurological disorders.

Stroke is a leading cause of death and the leading cause of long-term disability, but its electrophysiological basis is poorly understood. Characterizing acute ischemic neuronal activity dynamics is important for understanding the temporal and spatial development of ischemic pathophysiology and determining neuronal activity signatures of ischemia. Using a 32-microelectrode array spanning the depth of cortex, electrophysiological recordings generated for the first time a continuous spatiotemporal profile of local field potentials (LFP) and multi-unit activity (MUA) before (baseline) and directly after (0–5 h) distal, permanent MCA occlusion (pMCAo) in a rat model. Although evoked activity persisted for hours after pMCAo with minor differences from baseline, spatiotemporal analyses of spontaneous activity revealed that LFP became spatially and temporally synchronized regardless of cortical depth within minutes after pMCAo and extended over large parts of cortex. Such enhanced post-ischemic synchrony was found to be driven by increased bursts of low multi-frequency oscillations and continued throughout the acute ischemic period whereas synchrony measures minimally changed over the same recording period in surgical sham controls. EEG recordings of a similar frequency range have been applied to successfully predict stroke damage and recovery, suggesting clear clinical relevance for our rat model.

Intrinsic signal optical imaging (ISOI) remains one of the most exciting functional imaging techniques for functional mapping. It is routinely employed for basic research and has also been slowly adopted in recent years in preclinical and clinical research.

A common class of methods for analyzing of multivariate time series, stationary and nonstationary, decomposes the observed series into latent sources. Methods such as principal compoment analysis (PCA), independent component analysis (ICA) and Stationary Subspace Analysis (SSA) assume the observed multivariate process is generated by latent sources that are stationary or nonstationary. We develop a method that tracks changes in the complexity of a 32-channel local field potential (LFP) signal from a rat following an experimentally induced stroke. We study complexity through the latent sources and their dimensions that can change across epochs due to an induced shock to the cortical system. Our method compares the spread of spectral information in several multivariate stationary processes with different dimensions. A frequency specific spectral ratio (FS-ratio) statistic is proposed and its asymptotic properties are derived. The FS-ratio is blind to the dimension of the stationary process and captures the proportion of spectral information in various (user-specified) frequency bands. We apply our method to study differences in complexity and structure of the LFP before and after system shock. The analysis indicates that spectral information in the beta frequency band (12-30 Hertz) demonstrated the greatest change in structure and complexity due to the stroke., Comment: 34 pages

This paper intends to develop tools for characterizing non-linear spectral dependence between spontaneous brain signals. We use parametric copula models (both bivariate and vine models) applied on the magnitude of Fourier coefficients rather than using coherence. The motivation behind this work is an experiment on rats that studied the impact of stroke on the connectivity structure (dependence) between local field potentials recorded at various channels. We address the following major questions. First, we ask whether one can detect any changepoint in the regime of a brain channel for a given frequency band based on a difference between the cumulative distribution functions modeled for each epoch (small window of time). Our proposed approach is an iterative algorithm which compares each successive bivariate copulas on all the epochs range, using a bivariate Kolmogorov-Smirnov statistic. Second, we ask whether stroke can alter the dependence structure of brain signals; and examine whether changes in dependence are present only in some channels or generalized across channels. These questions are addressed by comparing Vine-copulas models fitted for each epoch. We provide the necessary framework and show the effectiveness of our methods through the results for the local field potential data analysis of a rat.

Employing wide-field optical imaging techniques supported by electrophysiological recordings, previous studies have demonstrated that stimulation of a spatially restricted area (point) in the sensory periphery results in a large evoked neuronal activity spread in mammalian primary cortices. In rats’ primary cortices, such large evoked spreads extend diffusely in multiple directions, cross cortical cytoarchitectural borders and can trespass into other unimodal sensory areas. These point spreads are supported by a spatially matching, diffuse set of long-range horizontal projections within gray matter that extend in multiple directions and cross borders to interconnect different cortical areas. This horizontal projection system is in addition to well-known area-to-area clustered projections to defined targets through white matter. Could similar two-projection cortical systems also be found in cortical regions that differ in their cytoarchitectural structure? To address this question, an adeno-associated viral vector expressing green fluorescent protein (GFP) was injected as an anterograde tract tracer into granular somatosensory cortex (trunk area), dysgranular cortex (somatosensory dysgranular zone and extrastriate cortex) and agranular motor cortex (MCx). Irrespective of the injection site the same two projection systems were found, and their quantification revealed a close similarity to findings in primary sensory cortices. Following detailed reconstruction, the diffuse horizontal axon radiation was found to possess numerous varicosities and to include short, medium and long axons, the latter extending up to 5.2 mm. These “proof of concept” findings suggest that the similarity of the two projection systems among different cortical areas could potentially constitute a canonical motif of neocortical organization.

© 2015, Springer-Verlag Berlin Heidelberg.Stimulation of a single whisker evokes a peak of activity that is centered over the associated barrel in rat primary somatosensory cortex, and yet the evoked local field potential and the intrinsic signal optical imaging response spread symmetrically away from this barrel for over 3.5 mm to cross cytoarchitectonic borders into other “unimodal” sensory cortical areas. To determine whether long horizontal axons have the spatial distribution necessary to underlie this activity spread, we injected adeno-associated viral vectors into barrel cortex and characterized labeled axons extending from the injection site in transverse sections of flattened cortex. Combined qualitative and quantitative analyses revealed labeled axons radiating diffusely in all directions for over 3.5 mm from supragranular injection sites, with density declining over distance. The projection pattern was similar at four different cortical depths, including infragranular laminae. Infragranular vector injections produced patterns similar to the supragranular injections. Long horizontal axons were detected both using a vector with a permissive cytomegalovirus promoter to label all neuronal subtypes and using a calcium/calmodulin-dependent protein kinase II α vector to restrict labeling to excitatory cortical pyramidal neurons. Individual axons were successfully reconstructed from series of supragranular sections, indicating that they traversed gray matter only. Reconstructed axons extended from the injection site, left the barrel field, branched, and sometimes crossed into other sensory cortices identified by cytochrome oxidase staining. Thus, radiations of long horizontal axons indeed have the spatial characteristics necessary to explain horizontal activity spreads. These axons may contribute to multimodal cortical responses and various forms of cortical neural plasticity.

This guest editorial summarizes Pioneers in Neurophotonics: Special Section Honoring Professor Amiram Grinvald.

This review brings together a collection of studies that specifically use wide-field high-resolution mesoscopic level imaging techniques (intrinsic signal optical imaging; voltage-sensitive dye optical imaging) to image the cortical point spread (PS): the total spread of cortical activation comprising a large neuronal ensemble evoked by spatially restricted (point) stimulation of the sensory periphery (e.g., whisker, pure tone, point visual stimulation). The collective imaging findings, combined with supporting anatomical and electrophysiological findings, revealed some key aspects about the PS including its very large (radius of several mm) and relatively symmetrical spatial extent capable of crossing cytoarchitectural borders and trespassing into other cortical areas; its relationship with underlying evoked subthreshold activity and underlying anatomical system of long-range horizontal projections within gray matter, both also crossing borders; its contextual modulation and plasticity; the ability of its relative spatiotemporal profile to remain invariant to major changes in stimulation parameters; its potential role as a building block for integrative cortical activity; and its ubiquitous presence across various cortical areas and across mammalian species. Together, these findings advance our understanding about the neocortex at the mesoscopic level by underscoring that the cortical PS constitutes a fundamental motif of neocortical structure-function relationship.

Using a rodent model of ischemic stroke (permanent middle cerebral artery occlusion; pMCAO), our laboratory has previously demonstrated that sensory-evoked cortical activation via mechanical single whisker stimulation treatment delivered under an anesthetized condition within 2 hours of ischemic onset confers complete protection from impending infarct. There is a limited time window for this protection: rats that received the identical treatment 3 hours following ischemic onset lost neuronal function and sustained a substantial infarct. Rats in these studies, however, were anesthetized with sodium pentobarbital or isoflurane, whereas most human stroke patients are typically awake. To optimize our animal model, the present study examined, using functional imaging, histological, and behavioral analysis, whether self-induced sensory-motor stimulation is also protective in unrestrained, behaving, rats that actively explore an enriched environment. Rats were revived from anesthesia either immediately or three hours after pMCAO – at which point they were allowed to freely explore an enriched environment. Rats that explored immediately after ischemic onset maintained normal cortical function and did not sustain infarct, even when their whiskers were clipped. Rats that were revived 3 hours post-pMCAO exhibited eliminated cortical function and sustained cortical infarct. Further, the data suggest that the level of individual active exploration could influence the outcome. Thus, early activation of the ischemic cortical area via unrestrained exploration results in protection from ischemic infarct, whereas late activation results in infarct, irrespective of level of arousal or whisker-specific stimulation.

There is a growing recognition regarding the importance of pial collateral flow in the protection from impending ischemic stroke both in preclinical and clinical studies. Collateral flow is also a major player in sensory stimulation-based protection from impending ischemic stroke. Doppler optical coherence tomography has been employed to image spatiotemporal patterns of collateral flow within the dorsal branches of the middle cerebral artery (MCA) as it provides a powerful tool for quantitative in vivo flow parameters imaging (velocity, flux, direction of flow, and radius of imaged branches). It was employed prior to and following dorsal permanent MCA occlusion (pMCAo) in rat models of treatment by protective sensory stimulation, untreated controls, or sham surgery controls. Unexpectedly, following pMCAo in the majority of subjects, some MCA branches continued to show anterograde blood flow patterns over time despite severing of the MCA. Further, in the presence of protective sensory stimulation, the anterograde velocity and flux were stronger and lasted longer than in retrograde flow branches, even within different branches of single subjects, but stimulated retrograde branches showed stronger flow parameters at 24 h. Our study suggests that the spatiotemporal patterns of collateral-based dorsal MCA flow are dynamic and provide a detailed description on the differential effects of protective sensory stimulation.

Using a rodent model of ischemia [permanent middle cerebral artery occlusion (pMCAO)], previous studies demonstrated that whisker stimulation treatment completely protects the cortex from impending stroke when initiated within 2 h following pMCAO. When initiated 3 h post-pMCAO, the identical treatment exacerbates stroke damage. Rats in these studies, however, were anesthetised with sodium pentobarbital, whereas human stroke patients are typically awake. To overcome this drawback, our laboratory has begun to use the anesthetic isoflurane, which allows rats to rapidly recover from pMCAO within minutes, to test stimulation treatment in awake rats and to determine whether isoflurane has an effect upon the pMCAO stroke model. We found no difference in infarct volume between pMCAO in untreated controls under either sodium pentobarbital or isoflurane, and the primary finding was that rats that received treatment immediately post-pMCAO maintain cortical function and no stroke damage, whereas rats that received treatment 3 h post-pMCAO exhibited eliminated cortical activity and extensive stroke damage. The only difference between anesthetics was the broad extent of evoked cortical activity observed during both functional imaging and electrophysiological recording, suggesting that the extent of evoked activity evident under isoflurane anesthesia is supported by underlying neuronal activity. Given the high degree of similarity with previous data, we conclude that the pMCAO stroke model is upheld with the use of isoflurane. This study demonstrated that the isoflurane-anesthetised rat pMCAO model can be used for cerebrovascular studies, and allows for highly detailed investigation of potential novel treatments for ischemic stroke using awake, behaving animals.

© 2017 SPIE. Doppler optical coherence tomography (DOCT) is considered one of the most promising functional imaging modalities for neuro biology research and has demonstrated the ability to quantify cerebral blood flow velocity at a high accuracy. However, the measurement of total absolute blood flow velocity (BFV) of major cerebral arteries is still a difficult problem since it not only relates to the properties of the laser and the scattering particles, but also relates to the geometry of both directions of the laser beam and the flow. In this paper, focusing on the analysis of cerebral hemodynamics, we presents a method to quantify the total absolute blood flow velocity in middle cerebral artery (MCA) based on volumetric vessel reconstruction from pure DOCT images. A modified region growing segmentation method is first used to localize the MCA on successive DOCT B-scan images. Vessel skeletonization, followed by an averaging gradient angle calculation method, is then carried out to obtain Doppler angles along the entire MCA. Once the Doppler angles are determined, the absolute blood flow velocity of each position on the MCA is easily found. Given a seed point position on the MCA, our approach could achieve automatic quantification of the fully distributed absolute BFV. Based on experiments conducted using a swept-source optical coherence tomography system, our approach could achieve automatic quantification of the fully distributed absolute BFV across different vessel branches in the rodent brain.

Stroke is the fourth leading cause of death in the United States and the leading cause of long-term disability. Ischemic stroke, due to an interruption in blood supply, is particularly prevalent; 87% of all strokes are ischemic. Unfortunately, current options for acute treatment are extremely limited and there is a great need for new treatment strategies. This review will discuss evidence that mild sensory stimulation can completely protect the jeopardized brain from an impending stroke in a rodent model. When delivered within the first 2 hours following ischemic onset, this stimulation results in complete protection, including a full reestablishment of cortical function, sensorimotor capabilities, and blood flow. Identical stimulation, however, initiated 3 hours following ischemic onset, results in an increase in damage compared with untreated animals. The protective effect is not specific to a single sensory modality, anesthesia, or age, and increasing evoked cortical activity by increasing stimulation accelerates recovery. Taken together, these findings demonstrate that cortical activity is a critical factor for protection and suggest a new, exciting potential avenue for the development of acute stroke treatment strategies that may produce a noninvasive, drug-free, equipment-free, and side effect–free means of protecting from ischemic stroke.

We have previously demonstrated that exposure of adult rat to a type of enriched environment, known as ‘naturalistic habitat’ (NH), induces extensive functional plasticity in the whiskers’ representations within the primary somatosensory cortex. Here we have investigated the molecular basis for such functional plasticity involved in this model. Based on the role of BDNF on synaptic plasticity and neuronal growth, the focus of this study is on BDNF and its downstream effectors CREB, synapsin I, and GAP-43. In particular, we determined the effects of natural whiskers use during 2, 7 or 28 days exposure to a NH on barrel cortex and hippocampus, as compared to standard cage controls. Naturalistic whiskers use resulted in increased levels of mRNAs and proteins for BDNF and its downstream effectors. Level changes for these markers were already detected after 2 days in the naturalistic habitat and grew larger over longer exposures (7 and 28 days). The cerebral cortex was found to be sensitive to the naturalistic habitat exposure at all time points, and more sensitive than the hippocampus to the trimming of the whiskers. Trimming of the whiskers decreased the level of most of the markers under study suggesting that whiskers exert a tonic influence on plasticity markers that can be further enhanced by naturalistic use. These results implicate BDNF and its downstream effectors in the plasticity induced by the naturalistic habitat. The critical action of experience on molecular substrates of plasticity seems to provide molecular basis for the design of experienced-based rehabilitative strategies to enhance brain function.

Background and Purpose— Using a rodent model of ischemia (permanent middle cerebral artery occlusion), our laboratory previously demonstrated that 4.27 minutes of patterned single-whisker stimulation delivered over 120 minutes can fully protect from impending damage when initiated within 2 hours of permanent middle cerebral artery occlusion (“early”). When initiated 3 hours postpermanent middle cerebral artery occlusion (“late”), stimulation resulted in irreversible damage. Here we investigate the effect of altering pattern, distribution, or amount of stimulation in this model. Methods— We assessed the cortex using functional imaging and histological analysis with altered stimulation treatment protocols. In 2 groups of animals we administered the same number of whisker deflections but in a random rather than patterned fashion distributed either over 120 minutes or condensed into 10 minutes postpermanent middle cerebral artery occlusion. We also tested increased (full-whisker array versus single-whisker) stimulation. Results— Early random whisker stimulation (condensed or dispersed) resulted in protection equivalent to early patterned stimulation. Early full-whisker array patterned stimulation also resulted in complete protection but promoted faster recovery. Late full-whisker array patterned stimulation, however, resulted in loss of evoked function and infarct volumes larger than those sustained by single-whisker counterparts. Conclusions— When induced early on after ischemic insult, stimulus-evoked cortical activity, irrespective of the parameters of peripheral stimulation that induced it, seems to be the important variable for neuroprotection.

Intrinsic Signal Optical Imaging (ISOI) can be used to map cortical function and organization. Because its detected signal lasts 10+ sec consisting of three phases, trials are typically collected using a long (tens of seconds) stimulus delivery interval (SDI) at the expense of efficiency, even when interested in mapping only the first signal phase (e.g., ISOI initial dip). It is unclear how the activity profile can change when stimuli are delivered at shorter intervals, and whether a short SDI can be implemented to improve efficiency. The goals of the present study are two-fold: characterize the ISOI activity profile when multiple stimuli are delivered at 4 sec intervals, and determine whether successful mapping can be attained from trials collected using an SDI of 4 sec (offering >10× increase in efficiency). Our results indicate that four stimuli delivered 4 sec apart evoke an activity profile different from the triphasic signal, consisting of signal dips in a series at the same frequency as the stimuli despite a strong rise in signal prior to the 2nd–4th stimuli. Visualization of such signal dips is dependent on using a baseline immediately prior to every stimulus. Use of the 4-sec SDI is confirmed to successfully map activity with a similar location in peak activity and increased areal extent and peak magnitude compared to using a long SDI. Additional experiments were performed to begin addressing issues such as SDI temporal jittering, response magnitude as a function of SDI duration, and application for successful mapping of cortical function topography.

Parcellation according to function (e.g., visual, somatosensory, auditory, motor) is considered a fundamental property of sensorimotor cortical organization, traditionally defined from cytoarchitectonics and mapping studies relying on peak evoked neuronal activity. In the adult rat, stimulation of single whiskers evokes peak activity at topographically appropriate locations within somatosensory cortex and provides an example of cortical functional specificity. Here, we show that single whisker stimulation also evokes symmetrical areas of suprathreshold and subthreshold neuronal activation that spread extensively away from peak activity, effectively ignoring cortical borders by spilling deeply into multiple cortical territories of different modalities (auditory, visual and motor), where they were blocked by localized neuronal activity blocker injections and thus ruled out as possibly caused by “volume conductance.” These symmetrical activity spreads were supported by underlying border-crossing, long-range horizontal connections as confirmed with transection experiments and injections of anterograde neuronal tracer experiments. We found such large evoked activation spreads and their underlying connections regardless of whisker identity, cortical layer, or axis of recorded responses, thereby revealing a large scale nonspecific organization of sensorimotor cortex based on a motif of large symmetrical activation spreads. Because the large activation spreads and their underlying horizontal connections ignore anatomical borders between cortical modalities, sensorimotor cortex could therefore be viewed as a continuous entity rather than a collection of discrete, delineated unimodal regions, an organization that could coexist with established specificity of cortical organization and that could serve as a substrate for associative learning, direct multimodal integration and recovery of function after injury.

© 2016 Optical Society of America. Doppler optical coherence tomography (DOCT) is considered one of the most promising functional imaging modalities for neuro biology research and has demonstrated the ability to quantify cerebral blood flow velocity at a high accuracy. However, the measurement of total absolute blood flow velocity (BFV) of major cerebral arteries is still a difficult problem since it is related to vessel geometry. In this paper, we present a volumetric vessel reconstruction approach that is capable of measuring the absolute BFV distributed along the entire middle cerebral artery (MCA) within a large field-of-view. The Doppler angle at each point of the MCA, representing the vessel geometry, is derived analytically by localizing the artery from pure DOCT images through vessel segmentation and skeletonization. Our approach could achieve automatic quantification of the fully distributed absolute BFV across different vessel branches. Experiments on rodents using swept-source optical coherence tomography showed that our approach was able to reveal the consequences of permanent MCA occlusion with absolute BFV measurement.

Recent advances in functional imaging and neuronal recording techniques demonstrate that the spatial spread and amplitude of whisker functional representation in the somatosensory cortex of the adult rodent is extensive, but subject to modulations. One of the strongest modulators is naturalistic whisker use. In the cortices of rodents that have been transferred from their home cage to live for an extensive period in a naturalistic habitat, there is suppression of evoked neuronal responses accompanied by contraction and sharpening of receptive fields, and contraction and weakening of whisker functional representations. These unexpected characteristics also describe modulations of whisker functional representations in the cortex of a freely exploring rodent during short whisker-based explorations. These and related findings suggest that cortical modulations and plasticity could follow a 'less is more' strategy and, therefore, highlight how different cortical strategies could be utilized for different behavioral demands.

Rats use their large facial whiskers to discriminate the spatial features of objects. Despite numerous electrophysiological recording studies in the central trigeminal whisker representations that document neurons tuned to the direction of whisker deflection, there is no behavioral evidence to date that rats can use their whiskers to discriminate between object orientations. In the present study, we characterized whisker-dependent orientation discrimination using a one-trial learning procedure. Sprague-Dawley rats were trained and tested in a three-arm 'Y-maze' that was outfitted with 180 independently moveable bars that protruded into the arms of the maze to contact the whiskers. On the first day, the maze was configured to have two arms with only horizontal bars and a third arm with only vertical bars and rats were allowed to freely explore all arms. On the second day, rats were isolated in one arm that contained only vertical bars as a conditioned stimulus (CS) and administered three mild foot shocks. On the third day, the maze was configured identically to the first day and rats were once again allowed to freely explore the maze. We measured the percentage of time spent in each arm of the maze and found that most rats spent significantly less time in the arm containing the CS after training compared to before training. Subsequent control experiments determined that the conditioned avoidance was attributable to orientation cues, was caused by the association of the conditioned and unconditioned stimulus and was whisker-dependent. Avoidance behavior was significantly reduced when the difference between the conditioned and non-conditioned orientation difference was reduced to 45 degrees. Thus, rats can discriminate object orientation with their whiskers and an estimate of their discrimination thresholds can be rapidly acquired through the application of a one-trial learning paradigm.

Much of what is known about the functional organization and plasticity of adult sensory cortex is derived from animals housed in standard laboratory cages. Here we report that the transfer of adult rats reared in standard laboratory cages to a naturalistic habitat modifies the functional and morphological organization of the facial whisker representation in the somatosensory 'barrel' cortex. Cortical whisker representations, visualized with repeated intrinsic signal optical imaging in the same animals, contracted by 46% after four to six weeks of exposure to the naturalistic habitat. Acute, multi-site extracellular recordings demonstrated suppressed evoked neuronal responses and smaller, sharper constituent receptive fields in the upper cortical layers (II/III), but not in the thalamic recipient layer (IV), of rats with naturalistic experience. Morphological plasticity of the layer IV barrel field was observed, but on a substantially smaller scale than the functional plasticity. Thus, transferring animals to an environment that promotes the expression of natural, innate behaviours induces a large-scale functional refinement of cortical sensory maps.

We have previously reported that topical application of nerve growth factor (NGF) to the barrel cortex of an adult rat rapidly augmented a whisker functional representation (WFR) by increasing its area and height within minutes after NGF application. In addition, we found that TrkA, the high-affinity NGF receptor, was only found on fibers projecting into the barrel cortex. Here we use a combination of techniques including chronic intrinsic signal optical imaging, neuronal fiber tracking and immunohistological techniques, to test the hypothesis that NGF-induced rapid cortical plasticity is mediated by the cortical projections of the basal forebrain cholinergic system (BFCS). Our studies localize the source of the cells in the BFCS that project to a single WFR and also demonstrate that TrkA-immunoreactive fibers in the cortex are also cholinergic and likely arise from the BFCS. In addition, by selectively lesioning the BFCS cortical fibers with the immunotoxin 192 IgG-saporin, we show that NGF-induced WFR-cortical plasticity is eliminated. These results, taken together with our previously reported imaging results that demonstrated that agonists of the cholinergic system (particularly nicotine) showed transient NGF-like augmentations of a WFR, implicate the BFCS cortical projections as necessary for NGF's rapid plasticity in the adult rat somatosensory cortex.

The molecular mechanisms that coordinate the functional organization of the mammalian neocortex are largely unknown. We tested the involvement of a putative guidance label, ephrin-A5, in the functional organization of the somatosensory cortex by quantifying the functional representations of individual whiskersin vivoin adult ephrin-A5 knock-out mice, using intrinsic signal optical imaging. In wild-type mice ephrin-A5 is expressed in a gradient in the somatosensory cortex during development. In adult ephrin-A5 knock-out mice, we found a spatial gradient of change in the amount of cortical territory shared by individual whisker functional representations across the somatosensory cortex, as well as a gradient of change in the distance between the functional representations. Both gradients of change were in correspondence with the developmental expression gradient of ephrin-A5 in wild-type mice. These changes involved malformations of the cortical spacing of the thalamocortical components, without concurrent malformations of the intracortical components of individual whisker functional representations. Overall, these results suggest that a developmental guidance label, such as ephrin-A5, is involved in establishing certain spatial relationships of the functional organization of the adult neocortex, and they underscore the advantage of investigating gene manipulation usingin vivofunctional imaging.

Optical Doppler tomography (ODT) is an imaging modality that takes advantage of the short coherence length of a broad-band light sources to perform micrometer-scale, cross-sectional imaging of tissue structure and blood flow dynamics simultaneously. The authors review in this paper the principal of ODT and its applications. Results from in vitro and in vivo model studies demonstrated that ODT can map the blood flow velocity profile with high spatial resolution in scattering medium. ODT detection mechanisms are illustrated using Monte Carlo simulations. The application of ODT to image brain hemodynamics is demonstrated. Finally, the authors discuss the limitations of the current technology and application of a phase resolved technique to improve image speed and quality.

Previous research from our lab has shown that when using a rodent model of ischemic stroke (permanent middle cerebral artery occlusion), mild sensory stimulation, when delivered within two hours of ischemic onset, completely protects the cortex from impending ischemic stroke damage when assessed 24 hours post-occlusion. However, the long-term stability of this protection remains unclear. Using intrinsic signal optical imaging for assessment of cortical function, laser speckle imaging for assessment of blood flow, a battery of behavioral tests and cresyl violet for histological assessment, the present study examined whether this protection was long-lasting. When assessed 4 months post-occlusion (this length of time being equivalent to 10-15 years in humans), rats receiving sensory stimulation treatment immediately after ischemic onset exhibit normal neuronal and vascular function, and they are behaviorally and histologically equivalent to healthy controls (surgical shams). Thus, the complete neuroprotection due to cortical activation via sensory stimulation remains stable with time. These findings add support to the translational potential of this sensory stimulation-based treatment.

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia). Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex, is the most common site of human stroke, and ischemia within its territory can result in extensive dysfunction or death. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA. Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

An important parameter often investigated in the characterization of cortical functional organization is the areal extent of functional modules. Because it allows the visualization of functional modules with high spatial resolution in a noninvasive way to the cortex, intrinsic signal optical imaging (ISI) can be employed for the quantification of these areal extents. The present paper describes the use of the normalized threshold analysis of areal extent quantification for the objective assessment of single-whisker functional representations in the primary somatosensory cortex of adult rats. As the success of areal extent quantification depends on the ability of ISI to allow visualization of cortical representations with minimal stimulus-dependent blood vessel representations, which are commonly encountered by ISI, the present paper also describes the further development of the intratrial analysis of visualization for minimizing these vessel representations. Both analyses are discussed with respect to their advantages as well as their inherent limitations.

The suprathreshold tonotopic organization of rat and guinea pig auditory cortex was investigated using intrinsic signal optical imaging through a thinned skull. Optical imaging revealed that suprathreshold pure sine wave tone stimulation (25-80 dB) evoked activity over large cortical areas that were tonotopically organized. Three-dimensional surface plots of the activated areas revealed "patchy' auditory-evoked activity consisting of numerous local peaks and valleys building to a maximum. Subsequent detailed electrophysiological mapping in the same subjects confirmed the localization of auditory-evoked activity based on optical imaging, including responses to a test frequency at cortical loci more than 2 octaves away from the threshold-defined isofrequency contour. The success of this technique in visualizing auditory cortex functional organization at suprathreshold stimulus levels will allow for future investigations of auditory cortex frequency representation, including representational plasticity induced by a variety of experimental manipulations.

The one-to-one relationship between whiskers, barrels, and barrel columns described for rat barrel cortex demonstrates that the organization of cortical function adheres to topographical and columnar principles. Supporting evidence is typically based on a single or few whiskers being stimulated, although behaving rats rely on the use of all their whiskers. Less is known about the cortical response when many whiskers are stimulated. Here, we use intrinsic signal optical imaging and supra- and sub-threshold electrophysiology recordings to map and characterize the cortical response to an array of all large whiskers. The cortical response was found to possess a single peak located centrally within a large activation spread, thereby no longer conveying information about the individual identities of the stimulated whiskers (e.g., many local peaks). Using modeling and pharmacological manipulations, we determined that this single central peak, plus other salient properties, can be predicted by and depends on large cortical activation spreads evoked by individual whisker stimulation. Compared to single whisker stimulation, the peak magnitude was comparable in strength and the response area was 2.6-fold larger, with both exhibiting a reduction in variability that was particularly pronounced (3.8x) for the peak magnitude. Findings extended to a different collection (subset) of whiskers. Our results indicate the rat barrel cortex response to multi-site stimulation transcends one-to-one topography to culminate in a large activation spread with a single central peak, and offer a potential neurobiological mechanism for the psychophysical phenomenon of multi-site stimulation being perceived as though a single, central site has been stimulated.

Background Accumulated research has shown that the older adult brain is significantly more vulnerable to stroke than the young adult brain. Although recent evidence in young adult rats demonstrates that single‐whisker stimulation can result in complete protection from ischemic damage after permanent middle cerebral artery occlusion ( pMCAO ), it remains unclear whether the same treatment would be effective in older animals. Methods and Results Aged rats (21 to 24 months of age) underwent pMCAO and subsequently were divided into “treated” and “untreated” groups. Treated aged rats received intermittent single‐whisker stimulation during a 120‐minute period immediately after pMCAO , whereas untreated aged rats did not. These animals were assessed using a battery of behavioral tests 1 week before and 1 week after pMCAO , after which their brains were stained for infarct. An additional treated aged group and a treated young adult group also were imaged with functional imaging. Results demonstrated that the recovery of treated aged animals was indistinguishable from that of the treated young adult animals. Treated aged rats had fully intact sensorimotor behavior and no infarct, whereas untreated aged rats were impaired and sustained cortical infarct. Conclusions Taken together, our results confirm that single‐whisker stimulation is protective in an aged rodent pMCAO model, despite age‐associated stroke vulnerability. These findings further suggest potential for translation to the more clinically relevant older adult human population.

We describe a technique that uses spatially modulated near-infrared NIR illumination to detect and map changes in both optical properties absorption and reduced scattering parameters and tissue composition oxy- and deoxyhemoglobin, total hemoglobin, and oxygen saturation during acute ischemic injury in the rat barrel cortex. Cerebral ischemia is induced using an open vascular occlu- sion technique of the middle cerebral artery MCA. Diffuse reflected NIR light 680 to 980 nm from the left parietal somatosensory cortex is detected by a CCD camera before and after MCA occlusion. Monte Carlo simulations are used to analyze the spatial frequency depen- dence of the reflected light to predict spatiotemporal changes in the distribution of tissue absorption and scattering properties in the brain. Experimental results from seven rats show a 17±4.7% increase in tissue concentration of deoxyhemoglobin and a 45±3.1, 23±5.4, and 21±2.2% decrease in oxyhemoglobin, total hemoglobin concen- tration and cerebral tissue oxygen saturation levels, respectively, 45 min following induction of cerebral ischemia. An ischemic index Iisch=ctHHb/ctO2Hb reveals an average of more then twofold con- trast after MCAo. The wavelength-dependence of the reduced scatter- ing i.e., scatter power decreased by 35±10.3% after MCA occlu- sion. Compared to conventional CCD-based intrinsic signal optical imaging ISOI, the use of structured illumination and model-based analysis allows for generation of separate maps of light absorption and scattering properties as well as tissue hemoglobin concentration. This potentially provides a powerful approach for quantitative monitoring and imaging of neurophysiology and metabolism with high spatiotem- poral resolution. © 2009 Society of Photo-Optical Instrumentation Engineers. DOI: 10.1117/1.3116709