Despite progress in reducing ischemic stroke damage, complete protection remains elusive. Here we demonstrate that, after permanent occlusion of a major cortical artery (middle cerebral artery; MCA), single whisker stimulation can induce complete protection of the adult rat cortex, but only if administered within a critical time window. Animals that receive early treatment are histologically and behaviorally equivalent to healthy controls and have normal neuronal function. Protection of the cortex clearly requires reperfusion to the ischemic area despite permanent occlusion. Using blood flow imaging and other techniques we found evidence of reversed blood flow into MCA branches from an alternate arterial source via collateral vessels (inter-arterial connections), a potential mechanism for reperfusion. These findings suggest that the cortex is capable of extensive blood flow reorganization and more importantly that mild sensory stimulation can provide complete protection from impending stroke given early intervention. Such non-invasive, non-pharmacological intervention has clear translational potential.

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.

Intrinsic signal optical imaging with red illumination (ISOI) is used extensively to provide high spatial resolution maps of stimulusevoked hemodynamic-related signals as an indirect means to map evoked neuronal activity. This evoked signal is generally described as beginning with an undershoot or "dip" in signal that is faster, more transient, and weaker compared with the subsequent signal overshoot. In contrast, the evoked signal detected with blood oxygen level-dependent (BOLD) functional magnetic resonance imaging (fMRI) is generally described as containing an undershoot after the initial dip and overshoot, even though it, too, detects hemodynamic-related signals and its first two phases appear complementary to those of ISOI. Here, we used ISOI with 635 nm illumination to image over 13.5 s after a 1 s stimulus delivery to detect and successfully use the ISOI undershoot phase for functional mapping. Eight spatiotemporal attributes were assessed per signal phase including maximum areal extent and peak magnitude, both of which were largest for the ISOI overshoot, followed by the undershoot and then the initial dip. Peak activity location did not colocalize well between the three phases; furthermore, we found mostly modest correlations between attributes within each phase and sparse correlations between phases. Extended (13.5 s) electrophysiology recordings did not exhibit a reoccurrence of evoked suprathreshold or subthreshold neuronal responses that could be associated with the undershoot. Beyond the undershoot, additional overshoot/undershoot fluctuations were also mapped, but were typically less spatiotemporally specific to stimulus delivery. Implications for ISOI and BOLD fMRI are discussed.

Damage or deprivation of a localized region of the skin surface has been shown to induce a selective expansion of adjacent skin surface representations in the adult somatosensory cortex. Here, we use repeated optical imaging in conjunction with single unit recordings to assess the plasticity of a single whisker's functional representation in the adult rat. We observed a large-scale expansion of a single whisker's functional representation following innocuous removal of all neighboring whiskers. Surprisingly, the same manipulation can also induce a large-scale contraction of the representation if the animal is removed from its home cage and given a brief opportunity to use its whiskers for active exploration of a different environment. Both the expansion and contraction reverse upon regrowth of the deprived whiskers. Thus, allowing the animal to use its deprived receptor organ in active exploration can determine the direction of plasticity in the adult cortex.

When delivered within 1 and in most cases 2 hours of permanent middle cerebral artery occlusion (pMCAO), mild sensory stimulation (intermittent single whisker stimulation) was shown to be completely neuroprotective according to assessment with multiple techniques 24 hours after pMCAO in a rodent model of ischemic stroke (Lay et al., 2010). The acute effect of stimulation treatment on the ischemic cortex however, had yet to be reported. Here we characterize cortical function and perfusion during the 120 minute whisker stimulation period in four experimental groups with treatment initiated 0, 1, 2 hours (protected groups) or 3 hours post-pMCAO (unprotected group) using multiple techniques. According to functional imaging, a gradual return of evoked whisker functional representation to baseline levels was initiated with treatment onset and completed within the treatment period. Evoked neuronal activity and reperfusion to the ischemic area also showed a gradual recovery in protected animals. Surprisingly, a similar recovery profile was observed in response to treatment in all protected animals, irrespective of treatment onset time. Non-stimulated pMCAO control group data demonstrate that reperfusion is not spontaneous. This makes the complete protection observed in the majority of animals stimulated at 2 hours post-pMCAO even more surprising as these animals recovered despite having been in this severely ischemic state for two full hours. In summary, when delivered within a 2 hour window post- pMCAO, whisker stimulation treatment initiated reperfusion and a gradual recovery of cortical function that was completed or nearly completed within the treatment period.

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.

The anatomical representations of the large facial whiskers, termed barrels, are topographically organized and highly segregated in the posteromedial barrel subfield (PMBSF) of rat layer IV primary somatosensory cortex. Although the functional representations of single whiskers are aligned with their appropriate barrels, their areal extents are rather large, spreading outward from the appropriate barrel along the tangential plane and thereby spanning multiple neighboring and non-neighboring barrels and septal regions. To date, single-whisker functional representations have been characterized primarily for whiskers whose corresponding barrels are located centrally within the PMBSF (central whiskers). Using intrinsic signal imaging verified with post-imaging single-unit recording, we demonstrate that border whiskers, whose barrels are located at the borders of the PMBSF, also evoke large activity areas that are similar in size to those of central whiskers but spread beyond the PMBSF and sometimes beyond primary somatosensory cortex into the neighboring dysgranular zones. This study indicates that the large functional representation of a single whisker is a basic functional feature of the rat whisker-to-barrel system and, combined with results from other studies, suggest that a large functional representation of a small, point-like area on the sensory epithelium may be a functional feature of primary sensory cortex in general.

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