Mesoscale functional connectivity in macaque visual areas.

Studies of resting-state functional connectivity (rsFC) have provided rich insights into the structures and functions of the human brain. However, most rsFC studies have focused on large-scale brain connectivity. To explore rsFC at a finer scale, we used intrinsic signal optical imaging to image the ongoing activity of the anesthetized macaque visual cortex. Differential signals from functional domains were used to quantify network-specific fluctuations. In 30-60 min resting-state imaging, a series of coherent activation patterns were observed in all three visual areas we examined (V1, V2, and V4). These patterns matched the known functional maps (ocular dominance, orientation, color) obtained in visual stimulation conditions. These functional connectivity (FC) networks fluctuated independently over time and exhibited similar temporal characteristics. Coherent fluctuations, however, were observed from orientation FC networks in different areas and even across two hemispheres. Thus, FC in the macaque visual cortex was fully mapped both on a fine scale and over a long range. Hemodynamic signals can be used to explore mesoscale rsFC in a submillimeter resolution.

Orientation anisotropies in macaque visual areas.

In mammals, a larger number of neurons in V1 are devoted to cardinal (horizontal and vertical) orientations than to oblique orientations. However, electrophysiological results from the macaque monkey visual cortex are controversial. Both isotropic and anisotropic orientation distributions have been reported. It is also unclear whether different visual areas along the visual hierarchy have different orientation anisotropies. We analyzed orientation maps in a large set of intrinsic signal optical imaging data and found that both V1 and V4 exhibited significant orientation anisotropies. However, their overrepresented orientations were very different: in V1, both cardinal and radial orientations were overrepresented, while in V4, only cardinal bias was presented. These findings suggest that different cortical areas have evolved to emphasize different features that are suitable for their functional purposes, a factor that needs to be considered when efforts are made to explain the relationships between the visual environment and the cortical representation and between the cortical representation and visual perception.

Function-specific projections from V2 to V4 in macaques.

Previous studies have revealed modular projections from area V2 to area V4 in macaques. Specifically, V2 neurons in cytochrome oxidase (CO)-rich thin and CO-sparse pale stripes project to distinct regions in V4. However, how these modular projections relate to the functional subcompartments of V4 remains unclear. In this study, we injected retrograde fluorescent tracers into V4 regions with different functional properties (color, orientation, and direction) that were identified with intrinsic signal optical imaging (ISOI). We examined the labeled neurons in area V2 and their locations with respect to the CO patterns. Covariation was observed between the functional properties of the V4 injection sites and the numbers of labeled neurons in particular CO stripes. This covariation indicates that the color domains in V4 mainly received inputs from thin stripes in V2, whereas V4 orientation domains received inputs from pale stripes. Although motion-sensitive domains are present in both V2 and V4, our results did not reveal a functional specific feedforward projection between them. These results confirmed previous findings of modular projections from V2 to V4 and provided functional specificity for these anatomical projections. Together, these findings indicate that color and form remain separate in ventral mid-level visual processing.

Processing of motion boundary orientation in macaque V2.

Human and nonhuman primates are good at identifying an object based on its motion, a task that is believed to be carried out by the ventral visual pathway. However, the neural mechanisms underlying such ability remains unclear. We trained macaque monkeys to do orientation discrimination for motion boundaries (MBs) and recorded neuronal response in area V2 with microelectrode arrays. We found 10.9% of V2 neurons exhibited robust orientation selectivity to MBs, and their responses correlated with monkeys' orientation-discrimination performances. Furthermore, the responses of V2 direction-selective neurons recorded at the same time showed correlated activity with MB neurons for particular MB stimuli, suggesting that these motion-sensitive neurons made specific functional contributions to MB discrimination tasks. Our findings support the view that V2 plays a critical role in MB analysis and may achieve this through a neural circuit within area V2.

Curvature-processing domains in primate V4.

Neurons in primate V4 exhibit various types of selectivity for contour shapes, including curves, angles, and simple shapes. How are these neurons organized in V4 remains unclear. Using intrinsic signal optical imaging and two-photon calcium imaging, we observed submillimeter functional domains in V4 that contained neurons preferring curved contours over rectilinear ones. These curvature domains had similar sizes and response amplitudes as orientation domains but tended to separate from these regions. Within the curvature domains, neurons that preferred circles or curve orientations clustered further into finer scale subdomains. Nevertheless, individual neurons also had a wide range of contour selectivity, and neighboring neurons exhibited a substantial diversity in shape tuning besides their common shape preferences. In strong contrast to V4, V1 and V2 did not have such contour-shape-related domains. These findings highlight the importance and complexity of curvature processing in visual object recognition and the key functional role of V4 in this process.

An Orientation Map for Disparity-Defined Edges in Area V4.

Binocular disparity information is an important source of 3D perception. Neurons sensitive to binocular disparity are found in almost all major visual areas in nonhuman primates. In area V4, disparity processes are suggested for the purposes of 3D-shape representation and fine disparity perception. However, whether neurons in V4 are sensitive to disparity-defined edges used in shape representation is not clear. Additionally, a functional organization for disparity edges has not been demonstrated so far. With intrinsic signal optical imaging, we studied functional organization for disparity edges in the monkey visual areas V1, V2, and V4. We found that there is an orientation map in V4 activated by edges purely defined by binocular disparity. This map is consistent with the orientation map obtained with regular luminance-defined edges, indicating a cue-invariant edge representation in this area. In contrast, such a map is much weaker in V2 and totally absent in V1. These findings reveal a hierarchical processing of 3D shape along the ventral pathway and the important role that V4 plays in shape-from-disparity detection.

Visual Motion Processing in Macaque V2.

In the primate visual system, direction-selective (DS) neurons are critical for visual motion perception. While DS neurons in the dorsal visual pathway have been well characterized, the response properties of DS neurons in other major visual areas are largely unexplored. Recent optical imaging studies in monkey visual cortex area 2 (V2) revealed clusters of DS neurons. This imaging method facilitates targeted recordings from these neurons. Using optical imaging and single-cell recording, we characterized detailed response properties of DS neurons in macaque V2. Compared with DS neurons in the dorsal areas (e.g., middle temporal area [MT]), V2 DS neurons have a smaller receptive field and a stronger antagonistic surround. They do not code speed or plaid motion but are sensitive to motion contrast. Our results suggest that V2 DS neurons play an important role in figure-ground segregation. The clusters of V2 DS neurons are likely specialized functional systems for detecting motion contrast.

Solving visual correspondence between the two eyes via domain-based population encoding in nonhuman primates.

Stereoscopic vision depends on correct matching of corresponding features between the two eyes. It is unclear where the brain solves this binocular correspondence problem. Although our visual system is able to make correct global matches, there are many possible false matches between any two images. Here, we use optical imaging data of binocular disparity response in the visual cortex of awake and anesthetized monkeys to demonstrate that the second visual cortical area (V2) is the first cortical stage that correctly discards false matches and robustly encodes correct matches. Our findings indicate that a key transformation for achieving depth perception lies in early stages of extrastriate visual cortex and is achieved by population coding.

Intrinsic signal optical imaging of visual brain activity: Tracking of fast cortical dynamics.

Hemodynamic-based brain imaging techniques are typically incapable of monitoring brain activity with both high spatial and high temporal resolutions. In this study, we have used intrinsic signal optical imaging (ISOI), a relatively high spatial resolution imaging technique, to examine the temporal resolution of the hemodynamic signal. We imaged V1 responses in anesthetized monkey to a moving light spot. Movies of cortical responses clearly revealed a focus of hemodynamic response traveling across the cortical surface. Importantly, at different locations along the cortical trajectory, response timecourses maintained a similar tri-phasic shape and shifted sequentially across cortex with a predictable delay. We calculated the time between distinguishable timecourses and found that the temporal resolution of the signal at which two events can be reliably distinguished is about 80 milliseconds. These results suggest that hemodynamic-based imaging is suitable for detecting ongoing cortical events at high spatial resolution and with temporal resolution relevant for behavioral studies.

Firing Frequency Maxima of Fast-Spiking Neurons in Human, Monkey, and Mouse Neocortex.

Cortical fast-spiking (FS) neurons generate high-frequency action potentials (APs) without apparent frequency accommodation, thus providing fast and precise inhibition. However, the maximal firing frequency that they can reach, particularly in primate neocortex, remains unclear. Here, by recording in human, monkey, and mouse neocortical slices, we revealed that FS neurons in human association cortices (mostly temporal) could generate APs at a maximal mean frequency (Fmean) of 338 Hz and a maximal instantaneous frequency (Finst) of 453 Hz, and they increase with age. The maximal firing frequency of FS neurons in the association cortices (frontal and temporal) of monkey was even higher (Fmean 450 Hz, Finst 611 Hz), whereas in the association cortex (entorhinal) of mouse it was much lower (Fmean 215 Hz, Finst 342 Hz). Moreover, FS neurons in mouse primary visual cortex (V1) could fire at higher frequencies (Fmean 415 Hz, Finst 582 Hz) than those in association cortex. We further validated our in vitro data by examining spikes of putative FS neurons in behaving monkey and mouse. Together, our results demonstrate that the maximal firing frequency of FS neurons varies between species and cortical areas.

Rivalry-Like Neural Activity in Primary Visual Cortex in Anesthetized Monkeys.

Two incongruent images viewed by the two eyes cause binocular rivalry, during which observers perceive continuous alternations between these two visual images. Previous studies in both humans and monkeys have shown that the primary visual cortex (V1) plays a critical role in the rivalry perception. However, it is unclear whether the rivalry activity observed in V1 relies on conscious influences. Here, we examine the responses of V1 in monkeys under general anesthesia. With intrinsic signal optical imaging and single-trial analysis, alternating activation of ocular dominance columns in V1 was observed during binocularly incongruent stimulation. Left- and right-eye columns exhibited counterphase activation, which were modulated by stimulus features in ways similar to those found in conscious human observers. These observations indicated that binocular rivalry occurs in V1 without consciousness, suggesting that the low-level automatic mechanisms play a more important role than previously believed in handling visual ambiguities. SIGNIFICANCE STATEMENT: When visual input is ambiguous, for example, in viewing bistable images, human subjects normally perceive one of the interpretations at a particular moment. Previous studies have shown that both low-level visual processing and high-level attention contribute to the establishment of the final visual perception. However, it is not clear whether attention is indispensable in such a process. Here we show that rivalry-like neural activity persisted in monkey V1 when the monkeys were anesthetized and viewed binocularly incongruent stimuli. Such activity has many key features similar to those observed in conscious human subjects. These findings indicate that low-level visual processes play a critical role in solving visual ambiguity such as binocular rivalry.

Spatial structure of neuronal receptive field in awake monkey secondary visual cortex (V2).

Visual processing depends critically on the receptive field (RF) properties of visual neurons. However, comprehensive characterization of RFs beyond the primary visual cortex (V1) remains a challenge. Here we report fine RF structures in secondary visual cortex (V2) of awake macaque monkeys, identified through a projection pursuit regression analysis of neuronal responses to natural images. We found that V2 RFs could be broadly classified as V1-like (typical Gabor-shaped subunits), ultralong (subunits with high aspect ratios), or complex-shaped (subunits with multiple oriented components). Furthermore, single-unit recordings from functional domains identified by intrinsic optical imaging showed that neurons with ultralong RFs were primarily localized within pale stripes, whereas neurons with complex-shaped RFs were more concentrated in thin stripes. Thus, by combining single-unit recording with optical imaging and a computational approach, we identified RF subunits underlying spatial feature selectivity of V2 neurons and demonstrated the functional organization of these RF properties.

An Orientation Map for Motion Boundaries in Macaque V2.

The ability to extract the shape of moving objects is fundamental to visual perception. However, where such computations are processed in the visual system is unknown. To address this question, we used intrinsic signal optical imaging in awake monkeys to examine cortical response to perceptual contours defined by motion contrast (motion boundaries, MBs). We found that MB stimuli elicit a robust orientation response in area V2. Orientation maps derived from subtraction of orthogonal MB stimuli aligned well with the orientation maps obtained with luminance gratings (LGs). In contrast, area V1 responded well to LGs, but exhibited a much weaker orientation response to MBs. We further show that V2 direction domains respond to motion contrast, which is required in the detection of MB in V2. These results suggest that V2 represents MB information, an important prerequisite for shape recognition and figure-ground segregation.

A motion direction preference map in monkey V4.

In the primate visual system, area V4 is located in the ventral pathway and is traditionally thought to be involved in processing color and form information. However, little is known about its functional role in processing motion information. Using intrinsic signal optical imaging over large fields of view in V1, V2, and V4, we mapped the direction of motion responses in anesthetized macaques. We found that V4 contains direction-preferring domains that are preferentially activated by stimuli moving in one direction. These direction-preferring domains normally occupy several restricted regions of V4 and tend to overlap with orientation- and color-preferring domains. Single-cell recordings targeting these direction-preferring domains also showed a clustering, as well as a columnar organization of V4 direction-selective neurons. These data suggest that, in contrast to the classical view, motion information is also processed in ventral pathway regions such as area V4.

Quantitative inference of population response properties across eccentricity from motion-induced maps in macaque V1.

Interpreting population responses in the primary visual cortex (V1) remains a challenge especially with the advent of techniques measuring activations of large cortical areas simultaneously with high precision. For successful interpretation, a quantitatively precise model prediction is of great importance. In this study, we investigate how accurate a spatiotemporal filter (STF) model predicts average response profiles to coherently drifting random dot motion obtained by optical imaging of intrinsic signals in V1 of anesthetized macaques. We establish that orientation difference maps, obtained by subtracting orthogonal axis-of-motion, invert with increasing drift speeds, consistent with the motion streak effect. Consistent with perception, the speed at which the map inverts (the critical speed) depends on cortical eccentricity and systematically increases from foveal to parafoveal. We report that critical speeds and response maps to drifting motion are excellently reproduced by the STF model. Our study thus suggests that the STF model is quantitatively accurate enough to be used as a first model of choice for interpreting responses obtained with intrinsic imaging methods in V1. We show further that this good quantitative correspondence opens the possibility to infer otherwise not easily accessible population receptive field properties from responses to complex stimuli, such as drifting random dot motions.

A motion direction map in macaque V2.

In mammals, the perception of motion starts with direction-selective neurons in the visual cortex. Despite numerous studies in monkey primary and second visual cortex (V1 and V2), there has been no evidence of direction maps in these areas. In the present study, we used optical imaging methods to study the organization of motion response in macaque V1 and V2. In contrast to the findings in other mammals (e.g., cats and ferrets), we found no direction maps in macaque V1. Robust direction maps, however, were found in V2 thick/pale stripes and avoided thin stripes. In many cases direction maps were located within thick stripes and exhibited pinwheel or linear organizations. The presence of motion maps in V2 points to a newfound prominence of V2 in motion processing, for contributing to motion perception in the dorsal pathway and/or for motion cue-dependent form perception in the ventral pathway.

Functional organization for color and orientation in macaque V4.

Visual area V4 in the macaque monkey is a cortical area that is strongly involved in color and shape perception. However, fundamental questions about V4 are still debated. V4 was initially characterized as a color-processing area, but subsequent studies revealed that it contains a diverse complement of cells, including those with preference for color, orientation, disparity and higher-order feature preferences. This has led to disputes and uncertainty about the role of V4 in vision. Using intrinsic signal optical imaging methods in awake, behaving monkeys, we found that different feature preferences are functionally organized in V4. Optical images revealed that regions with preferential response to color were largely separate from orientation-selective regions. Our results help to resolve long-standing controversies regarding functional diversity and retinotopy in V4 and indicate the presence of spatially biased distribution of featural representation in V4 in the ventral visual pathway.

Orientation and Direction-of-Motion Response in the Middle Temporal Visual Area (MT) of New World Owl Monkeys as Revealed by Intrinsic-Signal Optical Imaging.

Intrinsic-signal optical imaging was used to evaluate relationships of domains of neurons in middle temporal visual area (MT) selective for stimulus orientation and direction-of-motion. Maps of activation were elicited in MT of owl monkeys by gratings drifting back-and-forth, flashed stationary gratings and unidirectionally drifting fields of random dots. Drifting gratings, typically used to reveal orientation preference domains, contain a motion component that may be represented in MT. Consequently, this stimulus could activate groups of cells responsive to the motion of the grating, its orientation or a combination of both. Domains elicited from either moving or static gratings were remarkably similar, indicating that these groups of cells are responding to orientation, although they may also encode information about motion. To assess the relationship between domains defined by drifting oriented gratings and those responsive to direction-of-motion, the response to drifting fields of random dots was measured within domains defined from thresholded maps of activation elicited by the drifting gratings. The optical response elicited by drifting fields of random dots was maximal in a direction orthogonal to the map of orientation preference. Thus, neurons in domains selective for stimulus orientation are also selective for motion orthogonal to the preferred stimulus orientation.

The organization of orientation-selective, luminance-change and binocular- preference domains in the second (V2) and third (V3) visual areas of New World owl monkeys as revealed by intrinsic signal optical imaging.

Optical imaging was used to map patterns of visually evoked activation in the second (V2) and third (V3) visual areas of owl monkeys. Modular patterns of activation were produced in response to stimulation with oriented gratings, binocular versus monocular stimulation, and stimuli containing wide-field luminance changes. In V2, luminance-change domains tended to lie between domains selective for orientation. Regions preferentially activated by binocular stimulation co-registered with orientation-selective domains. Co-alignment of images with cytochrome oxidase (CO)-processed sections revealed functional correlates of 2 types of CO-dense regions in V2. Orientation-responsive domains and binocular domains were correlated with the locations of CO-thick stripes, and luminance-change domains were correlated with the locations of CO-thin stripes. In V3, orientation preference, luminance-change, and binocular preference domains were observed, but were more irregularly arranged than those in V2. Our data suggest that in owl monkey V2, consistent with that in macaque monkeys, modules for processing contours and binocularity exist in one type of compartment and that modules related to processing-surface features exist within a separate type of compartment.