Encoding luminance surfaces in the visual cortex of mice and monkeys: difference in responses to edge and center.

Luminance and spatial contrast provide information on the surfaces and edges of objects. We investigated neural responses to black and white surfaces in the primary visual cortex (V1) of mice and monkeys. Unlike primates that use their fovea to inspect objects with high acuity, mice lack a fovea and have low visual acuity. It thus remains unclear whether monkeys and mice share similar neural mechanisms to process surfaces. The animals were presented with white or black surfaces and the population responses were measured at high spatial and temporal resolution using voltage-sensitive dye imaging. In mice, the population response to the surface was not edge-dominated with a tendency to center-dominance, whereas in monkeys the response was edge-dominated with a "hole" in the center of the surface. The population response to the surfaces in both species exhibited suppression relative to a grating stimulus. These results reveal the differences in spatial patterns to luminance surfaces in the V1 of mice and monkeys and provide evidence for a shared suppression process relative to grating.

Microstimulation in the primary visual cortex: activity patterns and their relation to visual responses and evoked saccades.

Intracortical microstimulation (ICMS) in the primary visual cortex (V1) can generate the visual perception of a small point of light, termed phosphene, and evoke saccades directed to the receptive field of the stimulated neurons. Although ICMS is widely used, a direct measurement of the spatio-temporal patterns of neural activity evoked by ICMS and their relation to the neural responses evoked by visual stimuli or how they relate to ICMS-evoked saccades are still missing. To investigate this, we combined ICMS with voltage-sensitive dye imaging in V1 of behaving monkeys and measured neural activity at a high spatial (meso-scale) and temporal resolution. We then compared the population response evoked by small visual stimuli to those evoked by microstimulation. Both stimulation types evoked population activity that spread over few millimeters in V1 and propagated to extrastriate areas. However, the population responses evoked by ICMS have shown faster dynamics for the activation transients and the horizontal propagation of activity revealed a wave-like propagation. Finally, neural activity in the ICMS condition was higher for trials with evoked saccades as compared with trials without saccades. Our results uncover the spatio-temporal patterns evoked by ICMS and their relation to visual processing and saccade generation.

Computational modeling of color perception with biologically plausible spiking neural networks.

Biologically plausible computational modeling of visual perception has the potential to link high-level visual experiences to their underlying neurons' spiking dynamic. In this work, we propose a neuromorphic (brain-inspired) Spiking Neural Network (SNN)-driven model for the reconstruction of colorful images from retinal inputs. We compared our results to experimentally obtained V1 neuronal activity maps in a macaque monkey using voltage-sensitive dye imaging and used the model to demonstrate and critically explore color constancy, color assimilation, and ambiguous color perception. Our parametric implementation allows critical evaluation of visual phenomena in a single biologically plausible computational framework. It uses a parametrized combination of high and low pass image filtering and SNN-based filling-in Poisson processes to provide adequate color image perception while accounting for differences in individual perception.

Spatiotemporal patterns of population response in the visual cortex under isoflurane: from wakefulness to loss of consciousness.

Anesthetic drugs are widely used in medicine and research to mediate loss of consciousness (LOC). Isoflurane is a commonly used anesthetic drug; however, its effects on cortical sensory processing, in particular around LOC, are not well understood. Using voltage-sensitive dye imaging, we measured visually evoked neuronal population response from the visual cortex in awake and anesthetized mice at 3 increasing concentrations of isoflurane, thus controlling the level of anesthesia from wakefulness to deep anesthesia. At low concentration of isoflurane, the effects on neuronal measures were minor relative to the awake condition. These effects augmented with increasing isoflurane concentration, while around LOC point, they showed abrupt and nonlinear changes. At the network level, we found that isoflurane decreased the stimulus-evoked intra-areal spatial spread of local neural activation, previously reported to be mediated by horizontal connections, and also reduced intra-areal synchronization of neuronal population. The synchronization between different visual areas decreased with higher isoflurane levels. Isoflurane reduced the population response amplitude and prolonged their latencies while higher visual areas showed increased vulnerability to isoflurane concentration. Our results uncover the changes in neural activity and synchronization at isoflurane concentrations leading to LOC and suggest reverse hierarchical shutdown of cortical areas.

Advanced Circuit and Cellular Imaging Methods in Nonhuman Primates.

Novel genetically encoded tools and advanced microscopy methods have revolutionized neural circuit analyses in insects and rodents over the last two decades. Whereas numerous technical hurdles originally barred these methodologies from success in nonhuman primates (NHPs), current research has started to overcome those barriers. In some cases, methodological advances developed with NHPs have even surpassed their precursors. One such advance includes new ultra-large imaging windows on NHP cortex, which are larger than the entire rodent brain and allow analysis unprecedented ultra-large-scale circuits. NHP imaging chambers now remain patent for periods longer than a mouse's lifespan, allowing for long-term all-optical interrogation of identified circuits and neurons over timeframes that are relevant to human cognitive development. Here we present some recent imaging advances brought forth by research teams using macaques and marmosets. These include technical developments in optogenetics; voltage-, calcium- and glutamate-sensitive dye imaging; two-photon and wide-field optical imaging; viral delivery; and genetic expression of indicators and light-activated proteins that result in the visualization of tens of thousands of identified cortical neurons in NHPs. We describe a subset of the many recent advances in circuit and cellular imaging tools in NHPs focusing here primarily on the research presented during the corresponding mini-symposium at the 2019 Society for Neuroscience annual meeting.

Uncovering the Spatial Profile of Contour Integration from Fixational Saccades: Evidence for Widespread Processing in V1.

During contour integration, neuronal populations in the primary visual cortex (V1) enhance their responses to the contour while suppressing their responses to the noisy background. However, the spatial extent and profile of these responses are not fully understood. To investigate this question, 2 monkeys were trained on a contour detection task while we measured population responses in V1 using voltage-sensitive dyes. During stimulus presentation the animals made few fixational saccades, and we used their changing gaze position to image and analyze neuronal responses from large part of the stimulus, encoding multiple contour/background elements. We found that contour enhancement was present over the entire contour-mapped areas. The background suppression increased with distance from the contour, extending into background-mapped areas remotely located from the contour. The spatial profile of enhancement and suppression fitted well with a Gaussian model. These results imply that the divergent cortical responses to contour integration are modulated independently and extend over large areas in V1.

Population responses in V1 encode different figures by response amplitude.

The visual system simultaneously segregates between several objects presented in a visual scene. The neural code for encoding different objects or figures is not well understood. To study this question, we trained two monkeys to discriminate whether two elongated bars are either separate, thus generating two different figures, or connected, thus generating a single figure. Using voltage-sensitive dyes, we imaged at high spatial and temporal resolution V1 population responses evoked by the two bars, while keeping their local attributes similar among the two conditions. In the separate condition, unlike the connected condition, the population response to one bar is enhanced, whereas the response to the other is simultaneously suppressed. The response to the background remained unchanged between the two conditions. This divergent pattern developed ∼200 ms poststimulus onset and could discriminate well between the separate and connected single trials. The stimulus separation saliency and behavioral report were highly correlated with the differential response to the bars. In addition, the proximity and/or the specific location of the connectors seemed to have only a weak effect on this late activity pattern, further supporting the involvement of top-down influences. Additional neural codes were less informative about the separate and connected conditions, with much less consistency and discriminability compared with a response amplitude code. We suggest that V1 is involved in the encoding of each figure by different neuronal response amplitude, which can mediate their segregation and perception.

Representation of Color Surfaces in V1: Edge Enhancement and Unfilled Holes.

The neuronal mechanism underlying the representation of color surfaces in primary visual cortex (V1) is not well understood. We tested on color surfaces the previously proposed hypothesis that visual perception of uniform surfaces is mediated by an isomorphic, filled-in representation in V1. We used voltage-sensitive-dye imaging in fixating macaque monkeys to measure V1 population responses to spatially uniform chromatic (red, green, or blue) and achromatic (black or white) squares of different sizes (0.5°-8°) presented for 300 ms. Responses to both color and luminance squares early after stimulus onset were similarly edge-enhanced: for squares 1° and larger, regions corresponding to edges were activated much more than those corresponding to the center. At later times after stimulus onset, responses to achromatic squares' centers increased, partially "filling-in" the V1 representation of the center. The rising phase of the center response was slower for larger squares. Surprisingly, the responses to color squares behaved differently. For color squares of all sizes, responses remained edge-enhanced throughout the stimulus. There was no filling-in of the center. Our results imply that uniform filled-in representations of surfaces in V1 are not required for the perception of uniform surfaces and that chromatic and achromatic squares are represented differently in V1. SIGNIFICANCE STATEMENT: We used voltage-sensitive dye imaging from V1 of behaving monkeys to test the hypothesis that visual perception of uniform surfaces is mediated by an isomorphic, filled-in representation. We found that the early population responses to chromatic and achromatic surfaces are edge enhanced, emphasizing the importance of edges in surface processing. Next, we show for color surfaces that responses remained edge-enhanced throughout the stimulus presentation whereas response to luminance surfaces showed a slow neuronal 'filling-in' of the center. Our results suggest that isomorphic representation is not a general code for uniform surfaces in V1.

Reconstruction of shape contours from V1 activity at high resolution.

The role of primary visual cortex (V1) in encoding physical stimulus features is well known, while stimulus categorization is mainly attributed to higher visual areas. However, visual experience is not stripped down to invariant, categorical-only "labels." Rather, visual experiences are remarkably rich with details resulting in high-resolution perception of objects. If V1 is involved in this process, high-resolution readout of shape contours should be possible from V1 activity. To test this, we presented various shapes to awake, fixating monkeys while recording V1 population activity using voltage-sensitive dye imaging. A simplified bottom-up model was constructed based on known cortical properties and without an image prior. Contours were reconstructed from single trials, in sub-degree resolution by applying the inverse model to neuronal responses. These novel reconstruction results suggest V1 can be an important constituent in the detailed internal representation of visual experiences.

Figure-ground processing during fixational saccades in V1: indication for higher-order stability.

In a typical visual scene we continuously perceive a "figure" that is segregated from the surrounding "background" despite ongoing microsaccades and small saccades that are performed when attempting fixation (fixational saccades [FSs]). Previously reported neuronal correlates of figure-ground (FG) segregation in the primary visual cortex (V1) showed enhanced activity in the "figure" along with suppressed activity in the noisy "background." However, it is unknown how this FG modulation in V1 is affected by FSs. To investigate this question, we trained two monkeys to detect a contour embedded in a noisy background while simultaneously imaging V1 using voltage-sensitive dyes. During stimulus presentation, the monkeys typically performed 1-3 FSs, which displaced the contour over the retina. Using eye position and a 2D analytical model to map the stimulus onto V1, we were able to compute FG modulation before and after each FS. On the spatial cortical scale, we found that, after each FS, FG modulation follows the stimulus retinal displacement and "hops" within the V1 retinotopic map, suggesting visual instability. On the temporal scale, FG modulation is initiated in the new retinotopic position before it disappeared from the old retinotopic position. Moreover, the FG modulation developed faster after an FS, compared with after stimulus onset, which may contribute to visual stability of FG segregation, along the timeline of stimulus presentation. Therefore, despite spatial discontinuity of FG modulation in V1, the higher-order stability of FG modulation along time may enable our stable and continuous perception.

A contrast and surface code explains complex responses to black and white stimuli in V1.

We investigated the cortical mechanisms underlying the visual perception of luminance-defined surfaces and the preference for black over white stimuli in the macaque primary visual cortex, V1. We measured V1 population responses with voltage-sensitive dye imaging in fixating monkeys that were presented with white or black squares of equal contrast around a mid-gray. Regions corresponding to the squares' edges exhibited higher activity than those corresponding to the center. Responses to black were higher than to white, surprisingly to a much greater extent in the representation of the square's center. Additionally, the square-evoked activation patterns exhibited spatial modulations along the edges and corners. A model comprised of neural mechanisms that compute local contrast, local luminance temporal modulations in the black and white directions, and cortical center-surround interactions, could explain the observed population activity patterns in detail. The model captured the weaker contribution of V1 neurons that respond to positive (white) and negative (black) luminance surfaces, and the stronger contribution of V1 neurons that respond to edge contrast. Also, the model demonstrated how the response preference for black could be explained in terms of stronger surface-related activation to negative luminance modulation. The spatial modulations along the edges were accounted for by surround suppression. Overall the results reveal the relative strength of edge contrast and surface signals in the V1 response to visual objects.

Population response to natural images in the primary visual cortex encodes local stimulus attributes and perceptual processing.

The primary visual cortex (V1) is extensively studied with a large repertoire of stimuli, yet little is known about its encoding of natural images. Using voltage-sensitive dye imaging in behaving monkeys, we measured neural population response evoked in V1 by natural images presented during a face/scramble discrimination task. The population response showed two distinct phases of activity: an early phase that was spread over most of the imaged area, and a late phase that was spatially confined. To study the detailed relation between the stimulus and the population response, we used a simple encoding model to compute a continuous map of the expected neural response based on local attributes of the stimulus (luminance and contrast), followed by an analytical retinotopic transformation. Then, we computed the spatial correlation between the maps of the expected and observed response. We found that the early response was highly correlated with the local luminance of the stimulus and was sufficient to effectively discriminate between stimuli at the single trial level. The late response, on the other hand, showed a much lower correlation to the local luminance, was confined to central parts of the face images, and was highly correlated with the animal's perceptual report. Our study reveals a continuous spatial encoding of low- and high-level features of natural images in V1. The low level is directly linked to the stimulus basic local attributes and the high level is correlated with the perceptual outcome of the stimulus processing.

Spatiotemporal effects of microsaccades on population activity in the visual cortex of monkeys during fixation.

During visual fixation, the eyes make fast involuntary miniature movements known as microsaccades (MSs). When MSs are executed they displace the visual image over the retina and can generate neural modulation along the visual pathway. However, the effects of MSs on neural activity have substantial variability and are not fully understood. By utilizing voltage-sensitive dye imaging, we imaged the spatiotemporal patterns induced by MSs in V1 and V2 areas of behaving monkeys while they were fixating and presented with visual stimuli. We then investigated the neuronal modulation dynamics, induced by MSs, under different visual stimulation. MSs induced monophasic or biphasic neural responses depending on stimulus size. These neural responses were accompanied by different spatiotemporal patterns of synchronization. Finally, we show that a local patch of population response evoked by a small stimulus was clearly shifted over the V1 retinotopic map after each MS. Our results demonstrate the lack of visual stability in V1 following MSs and help clarify the substantial variability reported for MSs effects on neuronal responses. The observed neural effects suggest that MSs are associated with a continuum of neuronal responses in V1 area reflecting diverse spatiotemporal dynamics.

Spatio-temporal activation patterns of neuronal population evoked by optostimulation and the comparison to electrical microstimulation.

Optostimulation and electrical microstimulation are well-established techniques that enable to artificially stimulate the brain. While the activation patterns evoked by microstimulation in cortical network are well characterized, much less is known for optostimulation. Specifically, the activation maps of neuronal population at the membrane potential level and direct measurements of these maps were barely reported. In addition, only a few studies compared the activation patterns evoked by microstimulation and optostimulation. In this study we addressed these issues by applying optostimulation in the barrel cortex of anesthetized rats after a short (ShortExp) or a long (LongExp) opsin expression time and compared it to microstimulation. We measured the membrane potential of neuronal populations at high spatial (meso-scale) and temporal resolution using voltage-sensitive dye imaging. Longer optostimulation pulses evoked higher neural responses spreading over larger region relative to short pulses. Interestingly, similar optostimulation pulses evoked stronger and more prolonged population response in the LongExp vs. the ShortExp condition. Finally, the spatial activation patterns evoked in the LongExp condition showed an intermediate state, with higher resemblance to the microstimulation at the stimulation site. Therefore, short microstimulation and optostimulation can induce wide spread activation, however the effects of optostimulation depend on the opsin expression time.

Precise spatiotemporal patterns among visual cortical areas and their relation to visual stimulus processing.

Visual processing shows a highly distributed organization in which the presentation of a visual stimulus simultaneously activates neurons in multiple columns across several cortical areas. It has been suggested that precise spatiotemporal activity patterns within and across cortical areas play a key role in higher cognitive, motor, and visual functions. In the visual system, these patterns have been proposed to take part in binding stimulus features into a coherent object, i.e., to be involved in perceptual grouping. Using voltage-sensitive dye imaging (VSDI) in behaving monkeys (Macaca fascicularis, males), we simultaneously measured neural population activity in the primary visual cortex (V1) and extrastriate cortex (V2, V4) at high spatial and temporal resolution. We detected time point population events (PEs) in the VSDI signal of each pixel and found that they reflect transient increased neural activation within local populations by establishing their relation to spiking and local field potential activity. Then, we searched for repeating space and time relations between the detected PEs. We demonstrate the following: (1) spatiotemporal patterns occurring within (horizontal) and across (vertical) early visual areas repeat significantly above chance level; (2) information carried in only a few patterns can be used to reliably discriminate between stimulus categories on a single-trial level; (3) the spatiotemporal patterns yielding high classification performance are characterized by late temporal occurrence and top-down propagation, which are consistent with cortical mechanisms involving perceptual grouping. The pattern characteristics and the robust relation between the patterns and the stimulus categories suggest that spatiotemporal activity patterns play an important role in cortical mechanisms of higher visual processing.

Population response to contextual influences in the primary visual cortex.

Collinear proximal flankers can facilitate the detection of a low-contrast target or generate false-alarm target detection in the absence of a target. Although these effects are known to involve subthreshold neuronal interactions beyond the classical receptive field, the underlying neuronal mechanisms are not fully understood. Here, we used voltage-sensitive dye imaging that emphasizes subthreshold population activity, at high spatial and temporal resolution and imaged the visual cortex of fixating monkeys while they were presented with a low-contrast Gabor target, embedded within collinear or orthogonal flankers. We found that neuronal activity at the target site in area primary visual cortex increased and response latency decreased due to spatial spread of activation from the flankers' site. This increased activity was smaller than expected by a linear summation. The presentation of flankers alone induced strong spatial filling-in at the target site. Importantly, the increased neuronal activity at the target site was synchronized over time, both locally and with neuronal population at the flanker's site. This onset synchronization was higher for collinear than for orthogonal flankers. We further show that synchrony is a superior code over amplitude, for discriminating collinear from orthogonal pattern. These results suggest that population synchrony can serve as a code to discriminate contextual effects.

The effects of fixational eye movements on population responses in V1. Keynote at the 20th European Conference on Eye Movements in Alicante, September 22, 2019.

During visual fixation, the eyes make small and fast movements known as microsaccades (MSs). The effects of MSs on neural activity in the visual cortex are not well understood. Utilizing voltage-sensitive dye imaging, we imaged the spatiotemporal patterns of neuronal responses induced by MSs in early visual cortices of behaving monkeys. Our results reveal a continuous "visual instability" during fixation: while the visual stimulus moves over the retina with each MS, the neuronal activity in V1 'hops' within the retinotopic map, as dictated by the MS parameters. Neuronal modulations induced by MSs are characterized by neural suppression followed by neural enhancement and increased synchronization. The suppressed activity may underlie the suppressed perception during MSs whereas the late enhancement may facilitate the processing of new incoming image information. Moreover, the instability induced by MSs applies also to neural correlates of visual perception processes such as figure-ground (FG) segregation, which appear to develop faster after fixational saccades. Video stream: https://vimeo.com/362367119.

Columnar resolution of blood volume and oximetry functional maps in the behaving monkey; implications for FMRI.

The ultimate goal of high-resolution functional brain mapping is single-condition (stimulus versus no-stimulus maps) rather than differential imaging (comparing two "stimulus maps"), because the appropriate ("orthogonal") stimuli are rarely available. This requires some component(s) of activity-dependent hemodynamic signals to closely colocalize with electrical activity, like the early increase in deoxyhemoglobin, shown previously to yield high-quality functional single-condition maps. Conversely, nonlocal vascular responses dominate in cerebral blood volume (CBV)-based single-condition maps. Differential CBV maps are largely restricted to the parenchyma, implying that part of the CBV response does colocalize with electrical activity at fine spatial scale. By removing surface vascular activation from optical imaging data, we document the existence of a capillary CBV response component, regulated at fine spatial scale and yielding single-condition maps exhibiting approximately 100 microm resolution. Blood volume and -flow based single-condition functional mapping at columnar level should thus be feasible, provided that the capillary response component is selectively imaged.

Spatio-temporal characteristics of population responses evoked by microstimulation in the barrel cortex.

Intra-cortical microstimulation (ICMS) is a widely used technique to artificially stimulate cortical tissue. This method revealed functional maps and provided causal links between neuronal activity and cognitive, sensory or motor functions. The effects of ICMS on neural activity depend on stimulation parameters. Past studies investigated the effects of stimulation frequency mainly at the behavioral or motor level. Therefore the direct effect of frequency stimulation on the evoked spatio-temporal patterns of cortical activity is largely unknown. To study this question we used voltage-sensitive dye imaging to measure the population response in the barrel cortex of anesthetized rats evoked by high frequency stimulation (HFS), a lower frequency stimulation (LFS) of the same duration or a single pulse stimulation. We found that single pulse and short trains of ICMS induced cortical activity extending over few mm. HFS evoked a lower population response during the sustained response and showed a smaller activation across time and space compared with LFS. Finally the evoked population response started near the electrode site and spread horizontally at a propagation velocity in accordance with horizontal connections. In summary, HFS was less effective in cortical activation compared to LFS although HFS had 5 fold more energy than LFS.

Abnormal Population Responses in the Somatosensory Cortex of Alzheimer's Disease Model Mice.

Alzheimer's disease (AD) is the most common form of dementia. One of the neuropathological hallmarks of AD is the accumulation of amyloid-ß plaques. Overexpression of human amyloid precursor protein in transgenic mice induces hippocampal and neocortical amyloid-ß accumulation and plaque deposition that increases with age. The impact of these effects on neuronal population responses and network activity in sensory cortex is not well understood. We used Voltage Sensitive Dye Imaging, to investigate at high spatial and temporal resolution, the sensory evoked population responses in the barrel cortex of aged transgenic (Tg) mice and of age-matched non-transgenic littermate controls (Ctrl) mice. We found that a whisker deflection evoked abnormal sensory responses in the barrel cortex of Tg mice. The response amplitude and the spatial spread of the cortical responses were significantly larger in Tg than in Ctrl mice. At the network level, spontaneous activity was less synchronized over cortical space than in Ctrl mice, however synchronization during evoked responses induced by whisker deflection did not differ between the two groups. Thus, the presence of elevated Aß and plaques may alter population responses and disrupts neural synchronization in large-scale networks, leading to abnormalities in sensory processing.