Voltage-sensitive Dye Optical Imaging Research Articles

Behavioral Consequences of a Bifacial Map in the Mouse Somatosensory Cortex.

The whisker system is an important sensory organ with extensive neural representations in the brain of the mouse. Patterned neural modules (barrelettes) in the ipsilateral principal sensory nucleus of the trigeminal nerve (PrV) correspond to the whiskers. Axons of the PrV barrelette neurons cross the midline and confer the whisker-related patterning to the contralateral ventroposteromedial nucleus of the thalamus, and subsequently to the cortex. In this way, specific neural modules called barreloids and barrels in the contralateral thalamus and cortex represent each whisker. Partial midline crossing of the PrV axons, in a conditional Robo3 mutant (Robo3R3-5cKO) mouse line, leads to the formation of bilateral whisker maps in the ventroposteromedial, as well as the barrel cortex. We used voltage-sensitive dye optical imaging and somatosensory and motor behavioral tests to characterize the consequences of bifacial maps in the thalamocortical system. Voltage-sensitive dye optical imaging verified functional, bilateral whisker representation in the barrel cortex and activation of distinct cortical loci following ipsilateral and contralateral stimulation of the specific whiskers. The mutant animals were comparable with the control animals in sensorimotor tests. However, they showed noticeable deficits in all of the whisker-dependent or -related tests, including Y-maze exploration, horizontal surface approach, bridge crossing, gap crossing, texture discrimination, floating in water, and whisking laterality. Our results indicate that bifacial maps along the thalamocortical system do not offer a functional advantage. Instead, they lead to impairments, possibly due to the smaller size of the whisker-related modules and interference between the ipsilateral and contralateral whisker representations in the same thalamus and cortex.SIGNIFICANCE STATEMENT The whisker sensory system plays a quintessentially important role in exploratory behavior of mice and other nocturnal rodents. Here, we studied a novel mutant mouse line, in which the projections from the brainstem to the thalamus are disrupted. This led to formation of bilateral whisker maps in both the thalamus and the cortex. The two whisker maps crowd in a space normally devoted to the contralateral map alone and in a nonoverlapping fashion. Stimulation of the whiskers on either side activates the corresponding region of the map. Mice with bilateral whisker maps perform well in general sensorimotor tasks but show poor performance in specific tests that require whisker-dependent tactile discrimination. These observations indicate that contralateral, instead of bilateral, representation of the sensory space plays a critical role in acuity and fine discrimination during somesthesis.

Imaging Cajal's neuronal avalanche: how wide-field optical imaging of the point-spread advanced the understanding of neocortical structure-function relationship.

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.

Planar implantable sensor for in vivo measurement of cellular oxygen metabolism in brain tissue

BackgroundBrain imaging methods are continually improving. Imaging of the cerebral cortex is widely used in both animal experiments and charting human brain function in health and disease. Among the animal models, the rodent cerebral cortex has been widely used because of patterned neural representation of the whiskers on the snout and relative ease of activating cortical tissue with whisker stimulation. New methodWe tested a new planar solid-state oxygen sensor comprising a polymeric film with a phosphorescent oxygen-sensitive coating on the working side, to monitor dynamics of oxygen metabolism in the cerebral cortex following sensory stimulation. ResultsSensory stimulation led to changes in oxygenation and deoxygenation processes of activated areas in the barrel cortex. We demonstrate the possibility of dynamic mapping of relative changes in oxygenation in live mouse brain tissue with such a sensor. Comparison with existing methodOxygenation-based functional magnetic resonance imaging (fMRI) is very effective method for functional brain mapping but have high costs and limited spatial resolution. Optical imaging of intrinsic signal (IOS) does not provide the required sensitivity, and voltage-sensitive dye optical imaging (VSDi) has limited applicability due to significant toxicity of the voltage-sensitive dye. Our planar solid-state oxygen sensor imaging approach circumvents these limitations, providing a simple optical contrast agent with low toxicity and rapid application. ConclusionsThe planar solid-state oxygen sensor described here can be used as a tool in visualization and real-time analysis of sensory-evoked neural activity in vivo. Further, this approach allows visualization of local neural activity with high temporal and spatial resolution.

Study of the cortical representation of whisker frequency selectivity using voltage-sensitive dye optical imaging.

The facial whiskers of rodents act as a high-resolution tactile apparatus that allow the animal to detect the finest details of its environment. Previously it was shown that whisker-sensitive neurons in the somatosensory cortex show frequency selectivity to small amplitude stimuli, An intravital voltage-sensitive dye optical imaging (VSDi) method in combination with the different frequency whisker stimulation was used in order to visualize neural activity in the mice somatosensory cortex in response to the stimulation of a single whisker by different frequencies. Using the intravital voltage-sensitive dye optical imaging (VSDi) method in combination with the different frequency whisker stimulation we visualized neural activity in the mice somatosensory cortex in response to the stimulation of a single whisker by different frequencies. We found that whisker stimuli with different frequencies led to different optical signals in the barrel field. Our results provide evidence that different neurons of the barrel cortex have different frequency preferences. This supports prior research that whisker deflections cause responses in cortical neurons within the barrel field according to the frequency of the stimulation. Many studies of the whisker frequency selectivity were performed using unit recording but to map spatial organization, imaging methods are essential. In the work described in the present paper, we take a serious step toward detailed functional mapping of the somatosensory cortex using VSDi. To our knowledge, this is the first demonstration of whisker frequency sensitivity and selectivity of barrel cortex neurons with optical imaging methods.

Spatially Structured Sparse Morphological Component Separation for voltage-sensitive dye optical imaging

BackgroundVoltage-sensitive dye optical imaging is a promising technique for studying in vivo neural assemblies dynamics where functional clustering can be visualized in the imaging plane. Its practical potential is however limited by many artifacts. New methodWe present a novel method, that we call “SMCS” (Spatially Structured Sparse Morphological Component Separation), to separate the relevant biological signal from noise and artifacts. It extends Generalized Linear Models (GLM) by using a set of convex non-smooth regularization priors adapted to the morphology of the sources and artifacts to capture. ResultsWe make use of first order proximal splitting algorithms to solve the corresponding large scale optimization problem. We also propose an automatic parameters selection procedure based on statistical risk estimation methods. Comparison with existing methodsWe compare this method with blank subtraction and GLM methods on both synthetic and real data. It shows encouraging perspectives for the observation of complex cortical dynamics. ConclusionsThis work shows how recent advances in source separation can be integrated into a biophysical model of VSDOI. Going beyond GLM methods is important to capture transient cortical events such as propagating waves.

Study of the cortical representation of whisker directional deflection using voltage-sensitive dye optical imaging

Using voltage-sensitive dye optical imaging methods, we visualized neural activity in the rat barrel cortex in response to the deflection of a single whisker in different directions. Obtained data indicates that fast movements of single whiskers in varying directions correlate with different patterns of activation in the somatosensory cortex. A functional map was created based on the voltage-sensitive dye optical signal. This supports prior research that vibrissae deflections cause responses in different cortical neurons within the barrel field according to the direction of the deflection. By analogy with the orientation columns in the visual cortex, directionally biased single-whisker responses to different directions of deflection could be a possible mechanism for the directional selectivity of this important sensory response.

Whisker's Directional Selectivity: Orientation Columns in the Barrel Field?

AbstractUsing voltage-sensitive dye optical imaging methods, we visualized neural activity in the rat barrel cortex in response to the deflection of a single whisker in different directions. Obtained data indicates that fast movements of single whiskers in varying directions correlate with different patterns of activation in the somatosensory cortex. A functional map was created based on the voltage-sensitive dye optical signal. This supports prior research that vibrissae deflections cause responses in different cortical neurons within the barrel field according to the direction of the deflection. By analogy with the orientation columns in the visual cortex, directionally-biased single whisker responses to different directions of deflection could be a possible mechanism for the directional selectivity of this important sensory response.

Whisker's Directional Selectivity: Orientation Columns in the Barrel Field?

AbstractUsing voltage-sensitive dye optical imaging methods, we visualized neural activity in the rat barrel cortex in response to the deflection of a single whisker in different directions. Obtained data indicates that fast movements of single whiskers in varying directions correlate with different patterns of activation in the somatosensory cortex. A functional map was created based on the voltage-sensitive dye optical signal. This supports prior research that vibrissae deflections cause responses in different cortical neurons within the barrel field according to the direction of the deflection. By analogy with the orientation columns in the visual cortex, directionally-biased single whisker responses to different directions of deflection could be a possible mechanism for the directional selectivity of this important sensory response.

Membrane potential response profiles of CA1 pyramidal cells probed with voltage-sensitive dye optical imaging in rat hippocampal slices reveal the impact of GABA A-mediated feed-forward inhibition in signal propagation

The spatial and temporal distribution of excitatory and inhibitory membrane potential responses on a cell plays an important role in neuronal calculations in local neuronal circuits in the brain. The electrical dynamics of excitatory and inhibitory inputs along the somatodendritic extent of CA1 pyramidal cells during circuit activation were examined by stimulating strata radiatum (SR), oriens (SO), and lacunosum-moleculare (SLM) and measuring laminar responses with voltage-sensitive dye (VSD) optical recording methods. We first confirmed the linearity of the optical signal by comparing fluorescence changes in CA1 to global membrane potential changes when slices were bathed in high-potassium ([K +] O = 25 mM) solution. Except for a TTX-sensitive component in stratum pyramidale, fluorescence changes were equal in all strata, indicating that VSD sensitivity had reasonable linearity across layers. We then compared membrane potential profiles in slices exposed to picrotoxin, a GABA A receptor antagonist. We attributed the picrotoxin-induced changes in the first peak of the excitatory membrane potential to feed-forward inhibition and the later response (appearing 30 ms after stimulation) to feedback inhibition. A difference in feed-forward components was observed in perisomatic and distal apical dendritic regions after SR stimulation. SLM stimulation produced large differences in perisomatic and apical dendritic regions. SO stimulation, however, produced no feed-forward inhibition at the perisomatic region, but produces feed-forward inhibition in distal dendritic regions. These results suggest that actual inhibition of membrane potential response by feed-forward inhibition is greater at perisomatic regions after SR or SLM stimulation but is smaller at distal dendritic regions after SR, SO, and SLM stimulation.

Optical imaging of interaural time difference representation in rat auditory cortex

We used in vivo voltage-sensitive dye optical imaging to examine the cortical representation of interaural time difference (ITD), which is believed to be involved in sound source localization. We found that acoustic stimuli with dissimilar ITD activate various localized domains in the auditory cortex. The main loci of the activation pattern shift up to 1 mm during the first 40 ms of the response period. We suppose that some of the neurons in each pool are sensitive to the definite ITD and involved in the transduction of information about sound source localization, based on the ITD. This assumption gives a reasonable fit to the Jeffress model in which the neural network calculates the ITD to define the direction of the sound source. Such calculation forms the basis for the cortex's ability to detect the azimuth of the sound source.

1P208 膜電位感受性色素を用いて計測したラット海馬CA1錐体細胞におけるGABAシナプスを介したフィードフォワード抑制(神経回路・脳の情報処理,ポスター発表,第45回日本生物物理学会年会)

1P208 膜電位感受性色素を用いて計測したラット海馬CA1錐体細胞におけるGABAシナプスを介したフィードフォワード抑制(神経回路・脳の情報処理,ポスター発表,第45回日本生物物理学会年会)

Involvement of Kv1 Potassium Channels in Spreading Acidification and Depression in the Cerebellar Cortex

Spreading acidification and depression (SAD) is a form of propagated activity in the cerebellar cortex characterized by acidification and a transient depression in excitability. This study investigated the role of Kv1 potassium channels in SAD using neutral red, flavoprotein autofluorescence, and voltage-sensitive dye optical imaging in the mouse cerebellar cortex, in vivo. The probability of evoking SAD was greatly increased by blocking Kv1.1 as well as Kv1.2 potassium channels by their specific blockers dendrotoxin K (DTX-K) and tityustoxin (TsTX), respectively. DTX-K not only greatly lowered the threshold for evoking SAD but also resulted in multiple cycles of spread and spontaneous SAD. The occurrence of spontaneous SAD originating from spontaneous parallel fiber-like beams of activity suggests that blocking Kv1 channels increased parallel fiber excitability. This was confirmed by the generation of parallel fiber-like beams with the microinjection of glutamate into the upper molecular layer in the presence of DTX-K. The dramatic effects of DTX-K suggest a possible connection between SAD and episodic ataxia type 1 (EA1), a Kv1.1 potassium channelopathy. The threshold for evoking SAD was significantly lowered in the Kv1.1 heterozygous knockout mouse compared with wild-type littermates. Carbamazepine and acetazolamide, both effective in the treatment of EA1, significantly decreased the likelihood of evoking SAD. Blocking GABAergic neurotransmission did not alter the effectiveness of DTX-K. The cyclin D2 null mouse, which lacks cerebellar stellate cells, also exhibited SAD. Therefore blocking Kv1 potassium channels establishes the conditions needed to generate SAD. Furthermore, the results are consistent with the hypothesis that SAD may underlie the transient attacks of ataxia characterizing EA1.

Vitamin Content of Margarine

The whisker system is an important sensory organ with extensive neural representations in the brain of the mouse. Patterned neural modules (barrelettes) in the ipsilateral principal sensory nucleus of the trigeminal nerve (PrV) correspond to the whiskers. Axons of the PrV barrelette neurons cross the midline and confer the whisker-related patterning to the contralateral ventroposteromedial nucleus of the thalamus, and subsequently to the cortex. In this way, specific neural modules called barreloids and barrels in the contralateral thalamus and cortex represent each whisker. Partial midline crossing of the PrV axons, in a conditional Robo3 mutant (<i>Robo3^R3–5cKO</i>) mouse line, leads to the formation of bilateral whisker maps in the ventroposteromedial, as well as the barrel cortex. We used voltage-sensitive dye optical imaging and somatosensory and motor behavioral tests to characterize the consequences of bifacial maps in the thalamocortical system. Voltage-sensitive dye optical imaging verified functional, bilateral whisker representation in the barrel cortex and activation of distinct cortical loci following ipsilateral and contralateral stimulation of the specific whiskers. The mutant animals were comparable with the control animals in sensorimotor tests. However, they showed noticeable deficits in all of the whisker-dependent or -related tests, including Y-maze exploration, horizontal surface approach, bridge crossing, gap crossing, texture discrimination, floating in water, and whisking laterality. Our results indicate that bifacial maps along the thalamocortical system do not offer a functional advantage. Instead, they lead to impairments, possibly due to the smaller size of the whisker-related modules and interference between the ipsilateral and contralateral whisker representations in the same thalamus and cortex. <b>SIGNIFICANCE STATEMENT</b> The whisker sensory system plays a quintessentially important role in exploratory behavior of mice and other nocturnal rodents. Here, we studied a novel mutant mouse line, in which the projections from the brainstem to the thalamus are disrupted. This led to formation of bilateral whisker maps in both the thalamus and the cortex. The two whisker maps crowd in a space normally devoted to the contralateral map alone and in a nonoverlapping fashion. Stimulation of the whiskers on either side activates the corresponding region of the map. Mice with bilateral whisker maps perform well in general sensorimotor tasks but show poor performance in specific tests that require whisker-dependent tactile discrimination. These observations indicate that contralateral, instead of bilateral, representation of the sensory space plays a critical role in acuity and fine discrimination during somesthesis.