Long-term voltage-sensitive dye imaging reveals cortical dynamics in behaving monkeys

A novel method of chronic optical imaging based on new voltage-sensitive dyes (VSDs) was developed to facilitate the explorations of the spatial and temporal patterns underlying higher cognitive functions in the neocortex of behaving monkeys. Using this system, we were able to explore cortical dynamics, with high spatial and temporal resolution, over period of 相似文献   

The relationship between voltage-sensitive dye imaging signals and spiking activity of neural populations in primate V1

Voltage-sensitive dye imaging (VSDI) is a powerful technique for measuring neural population responses from a large cortical region simultaneously with millisecond temporal resolution and columnar spatial resolution. However, the relationship between the average VSDI signal and the average spiking activity of neural populations is largely unknown. To better understand this relationship, we compared visual responses measured from V1 of behaving monkeys using VSDI and single-unit electrophysiology. We found large and systematic differences between position and orientation tuning properties obtained with these two techniques. We then determined that a simple computational model could explain these tuning differences. This model, together with our experimental results, allowed us to estimate the quantitative relationship between the average VSDI signal and local spiking activity. We found that this relationship is similar to the previously reported nonlinear relationship between average membrane potential and spike rate in single V1 neurons, suggesting that VSDI signals are dominated by subthreshold synaptic activity. This model, together with the VSDI measured maps for spatial position (retinotopy) and orientation, also allowed us to estimate the spatial integration area over which neural responses contribute to the VSDI signal at a given location. We found that the VSDI-integration area is consistent with a Gaussian envelope with a space constant of ～230 μm. Finally, we show how this model and estimated parameters can be used to predict the pattern of population responses at the level of spiking activity from VSDI responses.     

Retinotopy and color sensitivity in human visual cortical area V8

Prior studies suggest the presence of a color-selective area in the inferior occipital-temporal region of human visual cortex. It has been proposed that this human area is homologous to macaque area V4, which is arguably color selective, but this has never been tested directly. To test this model, we compared the location of the human color-selective region to the retinotopic area boundaries in the same subjects, using functional magnetic resonance imaging (fMRI), cortical flattening and retinotopic mapping techniques. The human color-selective region did not match the location of area V4 (neither its dorsal nor ventral subdivisions), as extrapolated from macaque maps. Instead this region coincides with a new retinotopic area that we call 'V8', which includes a distinct representation of the fovea and both upper and lower visual fields. We also tested the response to stimuli that produce color afterimages and found that these stimuli, like real colors, caused preferential activation of V8 but not V4.     

Physiological correlates of perceptual learning in monkey V1 and V2

Performance in visual discrimination tasks improves with practice. Although the psychophysical parameters of these improvements have suggested the involvement of early areas in visual cortex, there has been little direct study of the physiological correlates of such perceptual learning at the level of individual neurons. To examine how neuronal response properties in the early visual system may change with practice, we trained monkeys for more than 6 mo in an orientation discrimination task in which behaviorally relevant stimuli were restricted to a particular retinal location and oriented around a specific orientation. During training the monkeys' discrimination thresholds gradually improved to much better than those of naive monkeys or humans. Although this improvement was specific to the trained orientation, it showed little retinotopic specificity. The receptive field properties of single neurons from regions representing the trained location and a location in the opposite visual hemifield were measured in V1 and V2. In most respects the receptive field properties in the representations of the trained and untrained regions were indistinguishable. However, in the regions of V1 and V2 representing the trained location, there were slightly fewer neurons whose optimal orientation was near the trained orientation. This resulted in a small but significant decrease in the V1 population response to the trained orientation at the trained location. Consequently, the observed neuronal populations did not exhibit any orientation-specific biases sufficient to explain the orientation specificity of the behavioral improvement. Pooling models suggest that the behavioral improvement was accomplished with a task-dependent and orientation-selective pooling of unaltered signals from early visual neurons. These data suggest that, even for training with stimuli suited to the selectivities found in early areas of visual cortex, behavioral improvements can occur in the absence of pronounced changes in the physiology of those areas.     

Cognitive mechanisms of transitive inference

Objects in natural scenes are rarely seen in isolation, but are usually overlapping or partially occluding other objects. To recognize individual objects, the visual system must be able to segregate overlapping objects from one another. Evidence from lesions in humans and monkeys suggest that perceptual segregation of occluded or overlapping objects involves extrastriate visual cortex. In monkeys, area V4 has been shown to play an important role in recognizing occluded or poorly salient shapes. In humans, a retinotopic homologue of ventral V4 (V4v) has been described, but it is not known whether this area is also functionally homologous to area V4 in monkeys. In this study, we tried to localize the visual cortical regions involved in perceptual segregation of overlapping shapes using positron emission tomography (PET). Regional cerebral blood flow (rCBF) was measured in seven subjects while they discriminated the relative areas of simultaneously presented rectangular shapes. In the control condition, the shapes were displayed without overlaps; in a second condition, the shapes overlapped each other partially. In a third condition, the shapes did not overlap but had been reduced in salience by adding random noise to the stimuli. Contrasting the overlapping shape condition with the control condition identified a single region in the left posterior lateral occipital cortex. The rCBF in this region also increased, though more weakly, during discrimination of shapes embedded in noise, relative to the control condition. The region activated by segregation of overlapping shapes was located in the posterior occipital cortex close to the anterior border of area V2, near the average location of human V4v as determined by retinotopic mapping studies. The activation of this region of extrastriate visual cortex by a task that involved segregation of overlapping shapes is consistent with monkey V4 and human V4v being functionally homologous. We conclude that discrimination of overlapping shapes involves in particular a region of extrastriate visual cortex located in the left lateral occipital cortex and that this region may correspond to human V4v.     

Visual responses of the human superior colliculus: a high-resolution functional magnetic resonance imaging study

The superior colliculus (SC) is a multimodal laminar structure located on the roof of the brain stem. The SC is a key structure in a distributed network of areas that mediate saccadic eye movements and shifts of attention across the visual field and has been extensively studied in nonhuman primates. In humans, it has proven difficult to study the SC with functional MRI (fMRI) because of its small size, deep location, and proximity to pulsating vascular structures. Here, we performed a series of high-resolution fMRI studies at 3 T to investigate basic visual response properties of the SC. The retinotopic organization of the SC was determined using the traveling wave method with flickering checkerboard stimuli presented at different polar angles and eccentricities. SC activations were confined to stimulation of the contralateral hemifield. Although a detailed retinotopic map was not observed, across subjects, the upper and lower visual fields were represented medially and laterally, respectively. Responses were dominantly evoked by stimuli presented along the horizontal meridian of the visual field. We also measured the sensitivity of the SC to luminance contrast, which has not been previously reported in primates. SC responses were nearly saturated by low contrast stimuli and showed only small response modulation with higher contrast stimuli, indicating high sensitivity to stimulus contrast. Responsiveness to stimulus motion in the SC was shown by robust activations evoked by moving versus static dot stimuli that could not be attributed to eye movements. The responses to contrast and motion stimuli were compared with those in the human lateral geniculate nucleus. Our results provide first insights into basic visual responses of the human SC and show the feasibility of studying subcortical structures using high-resolution fMRI.     

Eye position compensation improves estimates of response magnitude and receptive field geometry in alert monkeys

Studies of visual function in behaving subjects require that stimuli be positioned reliably on the retina in the presence of eye movements. Fixational eye movements scatter stimuli about the retina, inflating estimates of receptive field dimensions, reducing estimates of peak responses, and blurring maps of receptive field subregions. Scleral search coils are frequently used to measure eye position, but their utility for correcting the effects of fixational eye movements on receptive field maps has been questioned. Using eye coils sutured to the sclera and preamplifiers configured to minimize cable artifacts, we reexamined this issue in two rhesus monkeys. During repeated fixation trials, the eye position signal was used to adjust the stimulus position, compensating for eye movements and correcting the stimulus position to place it at the desired location on the retina. Estimates of response magnitudes and receptive field characteristics in V1 and in LGN were obtained in both compensated and uncompensated conditions. Receptive fields were narrower, with steeper borders, and response amplitudes were higher when eye movement compensation was used. In sum, compensating for eye movements facilitated more precise definition of the receptive field. We also monitored horizontal vergence over long sequences of fixation trials and found the variability to be low, as expected for this precise behavior. Our results imply that eye coil signals can be highly accurate and useful for optimizing visual physiology when rigorous precautions are observed.     

Functional organization of visual cortex in the prosimian bush baby revealed by optical imaging of intrinsic signals

Cells in primary visual cortex (V1) of primates and carnivores respond most strongly to a visual stimulus presented to one eye, in a particular visual field location, and at a particular orientation. Each of these stimulus attributes is mapped across the cortical surface, and, in macaque monkeys and cats, strong geometrical relationships exist between these feature maps. In macaque V1 and V2, correlations between feature maps and cytochrome oxidase (CO)-rich modules have also been observed. To see if such relationships reflect a conserved principle of V1 functional architecture among primate species, we examined these maps in the prosimian bush baby, a species that has been proposed to represent the ancestral primate organization. We found that the layout of individual feature maps in bush baby V1 is similar to that of other primates, but we found an entirely different organization of orientation preference in bush baby V2 compared with that reported in simian primates. Another striking distinction between bush baby and simian species is that we observed no strong relationships among maps of orientation, ocular dominance, and CO blobs in V1. Thus our findings suggest that precise relationships between feature maps are not a common element of the functional organization in all primates and that such relationships are not necessary for achieving basic coverage of stimulus feature combinations. In addition, our results suggest that specific relationships between feature maps in V1, and the subdivision of V2 into functional compartments, may have arisen comparatively late in the evolution of primates.     

Spatially specific FMRI repetition effects in human visual cortex

The functional MRI (fMRI) response to a pair of identical, successively presented stimuli can result in a smaller signal than the presentation of two nonidentical stimuli. This "repetition effect" has become a frequently used tool to make inferences about neural selectivity in specific cortical areas. However, little is known about the mechanism(s) underlying the effect. In particular, despite many successful applications of the technique in higher visual areas, repetition effects in lower visual areas [e.g., primary visual cortex (V1)] have been more difficult to characterize. One property that is well understood in early visual areas is the mapping of visual field locations to specific areas of the cortex (i.e., retinotopy). We used the retinotopic organization of V1 to activate progressively different populations of neurons in a rapid fMRI experimental design. We observed a repetition effect (reduced signal) when localized stimulus elements were repeated in identical locations. We show that this effect is spatially tuned and largely independent of both interstimulus interval (100-800 ms) and the focus of attention. Using the same timing parameters for which we observed a large effect of spatial position, we also examined the response to orientation changes and observed no effect of an orientation change on the response to repeated stimuli in V1 but significant effects in other retinotopic areas. Given these results, we discuss the possible causes of these repetition effects as well as the implications for interpreting other experiments that use this potentially powerful imaging technique.     

Functional imaging of the monkey brain.

Functional magnetic resonance imaging (fMRI) has become an essential tool for studying human brain function. Here we describe the application of this technique to anesthetized monkeys. We present spatially resolved functional images of the monkey cortex based on blood oxygenation level dependent (BOLD) contrast. Checkerboard patterns or pictures of primates were used to study stimulus-induced activation of the visual cortex, in a 4.7-Tesla magnetic field, using optimized multi-slice, gradient-recalled, echo-planar imaging (EPI) sequences to image the entire brain. Under our anesthesia protocol, visual stimulation yielded robust, reproducible, focal activation of the lateral geniculate nucleus (LGN), the primary visual area (V1) and a number of extrastriate visual areas, including areas in the superior temporal sulcus. Similar responses were obtained in alert, behaving monkeys performing a discrimination task.     

Relating retinotopic and object-selective responses in human lateral occipital cortex

What is the relationship between retinotopy and object selectivity in human lateral occipital (LO) cortex? We used functional magnetic resonance imaging (fMRI) to examine sensitivity to retinal position and category in LO, an object-selective region positioned posterior to MT along the lateral cortical surface. Six subjects participated in phase-encoded retinotopic mapping experiments as well as block-design experiments in which objects from six different categories were presented at six distinct positions in the visual field. We found substantial position modulation in LO using standard nonobject retinotopic mapping stimuli; this modulation extended beyond the boundaries of visual field maps LO-1 and LO-2. Further, LO showed a pronounced lower visual field bias: more LO voxels represented the lower contralateral visual field, and the mean LO response was higher to objects presented below fixation than above fixation. However, eccentricity effects produced by retinotopic mapping stimuli and objects differed. Whereas LO voxels preferred a range of eccentricities lying mostly outside the fovea in the retinotopic mapping experiment, LO responses were strongest to foveally presented objects. Finally, we found a stronger effect of position than category on both the mean LO response, as well as the distributed response across voxels. Overall these results demonstrate that retinal position exhibits strong effects on neural response in LO and indicates that these position effects may be explained by retinotopic organization.     

Neural responses in motor cortex and area 7a to real and apparent motion

The neural activity in area 7a and the arm area of motor cortex was recorded while real or path-guided apparent motion stimuli were presented to behaving monkeys in the absence of a motor response. A smooth stimulus motion was produced in the real motion condition, whereas in the apparent motion condition five stimuli were flashed successively at the vertices of a regular pentagon. The stimuli moved along a low contrast circular path with one of five speeds (180–540 deg/s). We found strong neural responses to real and apparent motion in area 7a and motor cortex. In the motor cortex, a substantial population of neurons showed a selective response to real moving stimuli in the absence of a motor response. This activity was modulated in some cases by the stimulus speed, and some of the neurons showed a response during a particular part of the circular trajectory of the stimulus; the preferred stimulus angular locations were evenly distributed across this neuronal ensemble. It is likely that these neural signals are continuously available to the motor cortex in order to generate responses that demand immediate action. In area 7a, two overlapping populations of neurons were observed. The first comprised cells the activity of which was tuned to the angular location of a circularly moving stimulus in the real motion condition. These cells also responded to apparent motion at high stimulus speeds. A visual receptive field analysis showed that the angular tuning in most of the area 7a neurons did not depend on the spatial location of the stimulus in relation to their receptive field. The second population was selective to apparent moving stimuli and showed a periodic entrainment of activation with the period of the inter-stimulus interval of the flashing dots. Both the angular location and the inter-stimulus interval neural signals can be used to generate precise behavioral responses towards real or apparent moving stimuli.     

Effect of spatial attention on the responses of area MT neurons

This study examines the influence of spatial attention on the responses of neurons in the middle temporal visual area (MT or V5) of extrastriate cortex. Two monkeys were trained to perform a direction-discrimination task. On each trial, two apertures of random-dot stimuli appeared simultaneously at two spatially separated locations; the monkeys were required to discriminate the direction of stimulus motion at one location while ignoring the stimulus at the other location. After extensive training, we recorded the responses of MT neurons in two configurations: 1) Both apertures placed "within" the neuron's receptive field (RF) and 2) one aperture covering the RF while the other was presented at a "remote" location. For each unit we compared the responses to identical stimulus displays when the monkey was instructed to attend to one or the other aperture. The responses of MT neurons were 8.7% stronger, on average, when the monkey attended to the spatial location that contained motion in the "preferred" direction. Attentional effects were equal, on average, in the within RF and remote configurations. The attentional modulations began approximately 300 ms after stimulus onset, gradually increased throughout the trial, and peaked near stimulus offset. An analysis of the neuronal responses on error trials suggests that the monkeys failed to attend to the appropriate spatial location on these trials. The relatively weak attentional effects that we observed contrast strikingly with recent results of Treue and Maunsell, who demonstrated very strong attentional modulations (median effect >80%) in MT in a task that shares many features with ours. Our results suggest that spatial attention alone is not sufficient to induce strong attentional effects in MT even when two competing motion stimuli appear within the RF of the recorded neuron. The difference between our results and those of Treue and Maunsell suggests that the magnitude of the attentional effects in MT may depend critically on how attention is directed to a particular stimulus and on the precise demands of the task.     

Two-dimensional mapping of the central and parafoveal visual field to human visual cortex

Primate visual cortex contains a set of maps of visual space. These maps are fundamental to early visual processing, yet their form is not fully understood in humans. This is especially true for the central and most important part of the visual field--the fovea. We used functional magnetic resonance imaging (fMRI) to measure the mapping geometry of human V1 and V2 down to 0.5 degrees of eccentricity. By applying automated atlas fitting procedures to parametrize and average retinotopic measurements of eight brains, we provide a reference standard for the two-dimensional geometry of human early visual cortex of unprecedented precision and analyze this high-quality mean dataset with respect to the 2-dimensional cortical magnification morphometry. The analysis indicates that 1) area V1 has meridional isotropy in areal projection: equal areas of visual space are mapped to equal areas of cortex at any given eccentricity. 2) V1 has a systematic pattern of local anisotropies: cortical magnification varies between isopolar and isoeccentricity lines, and 3) the shape of V1 deviates systematically from the complex-log model, the fit of which is particularly poor close to the fovea. We therefore propose that human V1 be fitted by models based on an equal-area principle of its two-dimensional magnification. 4) V2 is elongated by a factor of 2 in eccentricity direction relative to V1 and has significantly more local anisotropy. We propose that V2 has systematic intrinsic curvature, but V1 is intrinsically flat.     

No ON-OFF maps in supragranular layers of ferret visual cortex

Primary visual cortex contains functional maps of a number of stimulus properties including ocular dominance, orientation, direction, color, and spatial frequency. These maps must be organized with respect to each other and to a single continuous retinotopic map of visual space such that each stimulus parameter is represented at each point in space. In the ferret, geniculo-cortical inputs to cortical layer IV are segregated into ON- and OFF-center patches, suggesting the possibility that there might be an additional cortical map in this species. We have used optical imaging of intrinsic signals to search for ON-OFF maps in ferret visual cortical cells and have found none. This suggests that the high degree of ON-OFF segregation seen subcortically in the ferret may play a role in the development of visual cortical receptive fields rather than in adult cortical function.     

Top-down influence in early visual processing: a Bayesian perspective

Traditional views of visual processing suggest that early visual neurons are static spatiotemporal filters that extract local features by feedforward computation. The extracted information is then fed forward through a chain of modules to successively higher visual areas for further analysis. Recording from early visual neurons in awake behaving monkeys, we revealed there are many levels of complexity in the information processing of the early visual cortex. We found that the early visual neurons not only are sensitive to features within their receptive fields (RFs) but also to the global context of a visual scene, the behavioral relevance of the stimuli and the experience of the animals. These findings suggest that the early visual cortex (V1 and V2) is tightly coupled to and highly interactive with the rest of the visual system. The top-down interaction, mediated by recurrent feedback connections, introduces contextual information to influence the perceptual inference in the early visual cortex.     

Remapping in human visual cortex

With each eye movement, stationary objects in the world change position on the retina, yet we perceive the world as stable. Spatial updating, or remapping, is one neural mechanism by which the brain compensates for shifts in the retinal image caused by voluntary eye movements. Remapping of a visual representation is believed to arise from a widespread neural circuit including parietal and frontal cortex. The current experiment tests the hypothesis that extrastriate visual areas in human cortex have access to remapped spatial information. We tested this hypothesis using functional magnetic resonance imaging (fMRI). We first identified the borders of several occipital lobe visual areas using standard retinotopic techniques. We then tested subjects while they performed a single-step saccade task analogous to the task used in neurophysiological studies in monkeys, and two conditions that control for visual and motor effects. We analyzed the fMRI time series data with a nonlinear, fully Bayesian hierarchical statistical model. We identified remapping as activity in the single-step task that could not be attributed to purely visual or oculomotor effects. The strength of remapping was roughly monotonic with position in the visual hierarchy: remapped responses were largest in areas V3A and hV4 and smallest in V1 and V2. These results demonstrate that updated visual representations are present in cortical areas that are directly linked to visual perception.     

Precise alignment of micromachined electrode arrays with V1 functional maps

Recent theoretical models of primary visual cortex predict a relationship between receptive field properties and the location of the neuron within the orientation maps. Testing these predictions requires the development of new methods that allow the recording of single units at various locations across the orientation map. Here we present a novel technique for the precise alignment of functional maps and array recordings. Our strategy consists of first measuring the orientation maps in V1 using intrinsic optical imaging. A micromachined electrode array is subsequently implanted in the same patch of cortex for electrophysiological recordings, including the measurement of orientation tuning curves. The location of the array within the map is obtained by finding the position that maximizes the agreement between the preferred orientations measured electrically and optically. Experimental results of the alignment procedure from two implementations in monkey V1 are presented. The estimated accuracy of the procedure is evaluated using computer simulations. The methodology should prove useful in studying how signals from the local neighborhood of a neuron, thought to provide a dominant feedback signal, shape the receptive field properties in V1.     

Neural responses in the macaque v1 to bar stimuli with various lengths presented on the blind spot

Although there is no retinal input within the blind spot, it is filled with the same visual attributes as its surround. Earlier studies showed that neural responses are evoked at the retinotopic representation of the blind spot in the primary visual cortex (V1) when perceptual filling-in of a surface or completion of a bar occurs. To determine whether these neural responses correlate with perception, we recorded from V1 neurons whose receptive fields overlapped the blind spot. Bar stimuli of various lengths were presented at the blind spots of monkeys while they performed a fixation task. One end of the bar was fixed at a position outside the blind spot, and the position of the other end was varied. Perceived bar length was measured using a similar set of bar stimuli in human subjects. As long as one end of the bar was inside the blind spot, the perceived bar length remained constant, and when the bar exceeded the blind spot, perceptual completion occurred, and the perceived bar length increased substantially. Some V1 neurons of the monkey exhibited a significant increase in their activity when the bar exceeded the blind spot, even though the amount of the retinal stimulation increased only slightly. These response increases coincided with perceptual completion observed in human subjects and were much larger than would be expected from simple spatial summation and could not be explained by contextual modulation. We conclude that the completed bar appearing on the part of the receptive field embedded within the blind spot gave rise to the observed increase in neuronal activity.     

Dynamics of neuronal sensitivity in visual cortex and local feature discrimination

A striking aspect of natural scenes is that image features such as line orientation are strongly correlated at neighboring spatial locations but not at distant locations. Thus, during the viewing of a scene, eye movements are often accompanied by a change in the orientation structure of the image. How does this behavior influence the discrimination of local features and their encoding by visual cortical neurons? Here we examined the perceived changes in orientation induced by brief exposure to oriented image patterns in monkeys and humans, and then used reverse correlation to investigate dynamic changes in neuronal sensitivity in the primary visual cortex (V1) of behaving monkeys. Whereas brief adaptation to an oriented grating impaired identification of nearby orientations by broadening orientation selectivity and changing the preferred orientation of individual V1 neurons, it actually enhanced the identification of orthogonal orientations by sharpening neuronal selectivity. Hence, successive exposure to image patches of dissimilar spatial structure enhances both the ability to discriminate local features and the encoding of these features by V1 neurons.