Variable cardiac responses in rhesus macaque monkeys after discrete mediodorsal thalamus manipulations.

The control of some physiological parameters, such as the heart rate, is known to have a role in cognitive and emotional processes. Cardiac changes are also linked to mental health issues and neurodegeneration. Thus, it is not surprising that many of the brain structures typically associated with cognition and emotion also comprise a circuit-the central automatic network-responsible for the modulation of cardiovascular output. The mediodorsal thalamus (MD) is involved in higher cognitive processes and is also known to be connected to some of the key neural structures that regulate cardiovascular function. However, it is unclear whether the MD has any role in this circuitry. Here, we show that discrete manipulations (microstimulation during anaesthetized functional neuroimaging or localized cytotoxin infusions) to either the magnocellular or the parvocellular MD subdivisions led to observable and variable changes in the heart rate of female and male rhesus macaque monkeys. Considering the central positions that these two MD subdivisions have in frontal cortico-thalamocortical circuits, our findings suggest that MD contributions to autonomic regulation may interact with its identified role in higher cognitive processes, representing an important physiological link between cognition and emotion.

Structural Connectivity Changes After Fornix Transection in Macaques Using Probabilistic Diffusion Tractography.

The fornix, the limbic system's white matter tract connecting the extended hippocampal system to subcortical structures of the medial diencephalon, is strongly associated with learning and memory in humans and nonhuman primates (NHPs). Here, we sought to investigate alterations in structural connectivity across key cortical and subcortical regions after fornix transection in NHPs. We collected diffusion-weighted MRI (dMRI) data from three macaque monkeys that underwent bilateral fornix transection during neurosurgery and from four age- and cohort-matched control macaques that underwent surgery to implant a head-post but remained neurologically intact. dMRI data were collected from both groups at two time points, before and after the surgeries, and scans took place at around the same time for the two groups. We used probabilistic tractography and employed the number of tracking streamlines to quantify connectivity across our regions of interest (ROIs), in all dMRI sessions. In the neurologically intact monkeys, we observed high connectivity across certain ROIs, including the CA3 hippocampal subfield with the retrosplenial cortex (RSC), the anterior thalamus with the RSC, and the RSC with the anterior cingulate cortex (ACC). However, we found that, compared to the control group, the fornix-transected monkeys showed marked, significant, connectivity changes including increases between the anterior thalamus and the ACC and between the CA3 and the ACC, as well as decreases between the CA3 and the RSC. Our results highlight cortical and subcortical network changes after fornix transection and identify candidate indirect connectivity routes that may support memory functions after damage and/or neurodegeneration.

Single-Unit Recordings Reveal the Selectivity of a Human Face Area.

The exquisite capacity of primates to detect and recognize faces is crucial for social interactions. Although disentangling the neural basis of human face recognition remains a key goal in neuroscience, direct evidence at the single-neuron level is limited. We recorded from face-selective neurons in human visual cortex in a region characterized by functional magnetic resonance imaging (fMRI) activations for faces compared with objects. The majority of visually responsive neurons in this fMRI activation showed strong selectivity at short latencies for faces compared with objects. Feature-scrambled faces and face-like objects could also drive these neurons, suggesting that this region is not tightly tuned to the visual attributes that typically define whole human faces. These single-cell recordings within the human face processing system provide vital experimental evidence linking previous imaging studies in humans and invasive studies in animal models.SIGNIFICANCE STATEMENT We present the first recordings of face-selective neurons in or near an fMRI-defined patch in human visual cortex. Our unbiased multielectrode array recordings (i.e., no selection of neurons based on a search strategy) confirmed the validity of the BOLD contrast (faces-objects) in humans, a finding with implications for all human imaging studies. By presenting faces, feature-scrambled faces, and face-pareidolia (perceiving faces in inanimate objects) stimuli, we demonstrate that neurons at this level of the visual hierarchy are broadly tuned to the features of a face, independent of spatial configuration and low-level visual attributes.

Temporal dynamics of neural activity in macaque frontal cortex assessed with large-scale recordings.

The cortical network controlling the arm and hand when grasping objects consists of several areas in parietal and frontal cortex. Recently, more anterior prefrontal areas have also been implicated in object grasping, but their exact role is currently unclear. To investigate the neuronal encoding of objects during grasping in these prefrontal regions and their relation with other cortical areas of the grasping network, we performed large-scale recordings (more than 2000 responsive sites) in frontal cortex of monkeys during a saccade-reach-grasp task. When an object appeared in peripheral vision, the first burst of activity emerged in prearcuate areas (the FEF and area 45B), followed by dorsal and ventral premotor cortex, and a buildup of activity in primary motor cortex. After the saccade, prearcuate activity remained elevated while primary motor and premotor activity rose in anticipation of the upcoming arm and hand movement. Remarkably, a large number of premotor and prearcuate sites responded when the object appeared in peripheral vision and remained active when the object came into foveal vision. Thus, prearcuate and premotor areas continuously encode object information when directing gaze and grasping objects.

Effective Connectivity Reveals an Interconnected Inferotemporal Network for Three-Dimensional Structure Processing.

Disparity-defined 3D shape is processed in both the ventral and the dorsal visual stream. The network of cortical areas that is activated during the processing of disparity-defined 3D shape includes, in addition to parietal and premotor areas, three clearly distinct regions in inferotemporal cortex (ITC). To investigate the connectivity of the latter regions, we combined electrical stimulation with fMRI in male macaque monkeys. Electrical stimulation of each of the 3D-structure nodes in ITC mainly elicited increased fMRI activations in the other 3D-structure nodes and more variably in other parts of ventral visual cortex. Importantly, no increased activation was found in parietal areas, nor in PFC, whereas microstimulation in posterior parietal cortex did activate the ITC. Our results indicate that 3D-structure nodes in ITC form a strongly interconnected network, receiving input from parietal areas implicated in 3D-structure processing.SIGNIFICANCE STATEMENT Previous studies combining electrical microstimulation with functional imaging showed an interconnected set of regions in the ventral stream processing faces or bodies, but is has been unclear whether the same is true for other visual categories. Here the authors show that there is a connected system of stereo-selective regions in inferotemporal cortex, receiving input from parietal areas in the dorsal stream.

Corticocortical and Thalamocortical Changes in Functional Connectivity and White Matter Structural Integrity after Reward-Guided Learning of Visuospatial Discriminations in Rhesus Monkeys.

The frontal cortex and temporal lobes together regulate complex learning and memory capabilities. Here, we collected resting-state functional and diffusion-weighted MRI data before and after male rhesus macaque monkeys received extensive training to learn novel visuospatial discriminations (reward-guided learning). We found functional connectivity changes in orbitofrontal, ventromedial prefrontal, inferotemporal, entorhinal, retrosplenial, and anterior cingulate cortices, the subicular complex, and the dorsal, medial thalamus. These corticocortical and thalamocortical changes in functional connectivity were accompanied by related white matter structural alterations in the uncinate fasciculus, fornix, and ventral prefrontal tract: tracts that connect (sub)cortical networks and are implicated in learning and memory processes in monkeys and humans. After the well-trained monkeys received fornix transection, they were impaired in learning new visuospatial discriminations. In addition, the functional connectivity profile that was observed after the training was altered. These changes were accompanied by white matter changes in the ventral prefrontal tract, although the integrity of the uncinate fasciculus remained unchanged. Our experiments highlight the importance of different communication relayed among corticocortical and thalamocortical circuitry for the ability to learn new visuospatial associations (learning-to-learn) and to make reward-guided decisions.SIGNIFICANCE STATEMENT Frontal neural networks and the temporal lobes contribute to reward-guided learning in mammals. Here, we provide novel insight by showing that specific corticocortical and thalamocortical functional connectivity is altered after rhesus monkeys received extensive training to learn novel visuospatial discriminations. Contiguous white matter fiber pathways linking these gray matter structures, namely, the uncinate fasciculus, fornix, and ventral prefrontal tract, showed structural changes after completing training in the visuospatial task. Additionally, different patterns of functional and structural connectivity are reported after removal of subcortical connections within the extended hippocampal system, via fornix transection. These results highlight the importance of both corticocortical and thalamocortical interactions in reward-guided learning in the normal brain and identify brain structures important for memory capabilities after injury.

Localization of movable electrodes in a multi-electrode microdrive in nonhuman primates.

BACKGROUND: Recently, large-scale semi-chronic recording systems have been developed, unique in their capability to record simultaneously from multiple individually moveable electrodes. As these recording systems can cover a large area, knowledge of the exact location of each individual electrode is crucial. Currently, the only method of keeping track of electrode depth and thus location is through detailed notebook keeping on neural activity. NEW METHOD: We have improved the electrode localization by combining pre- and postoperative anatomical magnetic resonance imaging (MRI) scans with high resolution computed tomography (CT) scans throughout the experiment, and validated our method by comparing the resulting location estimates with traditional notebook-keeping. Finally, the actual location of a selection of electrodes was marked at the end of the experiment by creating small metallic depositions using electrical stimulation, and thereby made visible on MRI. RESULTS: Combining CT scans with a high resolution, artefact reducing sequence during the experiment with a preoperative MRI scan provides crucial information about the exact electrode location of multielectrode arrays with individually moveable electrodes. COMPARISON WITH EXISTING METHODS: The information obtained from the hybrid CT-MR image and the notes on spiking activity showed a similar pattern, with the clear advantage of the visualization of the exact position of the electrodes using our method. CONCLUSIONS: The described technique allows for a precise anatomical identification of the recorded brain areas and thus to draw strong conclusions about the role of each targeted cortical area in the behavior under study.

Correction: Single-cell selectivity and functional architecture of human lateral occipital complex.

[This corrects the article DOI: 10.1371/journal.pbio.3000280.].

Single-cell selectivity and functional architecture of human lateral occipital complex.

The human lateral occipital complex (LOC) is more strongly activated by images of objects compared to scrambled controls, but detailed information at the neuronal level is currently lacking. We recorded with microelectrode arrays in the LOC of 2 patients and obtained highly selective single-unit, multi-unit, and high-gamma responses to images of objects. Contrary to predictions derived from functional imaging studies, all neuronal properties indicated that the posterior subsector of LOC we recorded from occupies an unexpectedly high position in the hierarchy of visual areas. Notably, the response latencies of LOC neurons were long, the shape selectivity was spatially clustered, LOC receptive fields (RFs) were large and bilateral, and a number of LOC neurons exhibited three-dimensional (3D)-structure selectivity (a preference for convex or concave stimuli), which are all properties typical of end-stage ventral stream areas. Thus, our results challenge prevailing ideas about the position of the more posterior subsector of LOC in the hierarchy of visual areas.

Preserved extrastriate visual network in a monkey with substantial, naturally occurring damage to primary visual cortex.

Lesions of primary visual cortex (V1) lead to loss of conscious visual perception with significant impact on human patients. Understanding the neural consequences of such damage may aid the development of rehabilitation methods. In this rare case of a Rhesus macaque (monkey S), likely born without V1, the animal's in-group behaviour was unremarkable, but visual task training was impaired. With multi-modal magnetic resonance imaging, visual structures outside of the lesion appeared normal. Visual stimulation under anaesthesia with checkerboards activated lateral geniculate nucleus of monkey S, while full-field moving dots activated pulvinar. Visual cortical activation was sparse but included face patches. Consistently across lesion and control monkeys, functional connectivity analysis revealed an intact network of bilateral dorsal visual areas temporally correlated with V5/MT activation, even without V1. Despite robust subcortical responses to visual stimulation, we found little evidence for strengthened subcortical input to V5/MT supporting residual visual function or blindsight-like phenomena.

Effective chair training methods for neuroscience research involving rhesus macaques (Macaca mulatta).

BACKGROUND: Neuroscience research using macaques remains fundamental in our endeavours to understand how the human brain functions. Applying the refinement principle of the 3Rs is essential to optimise the monkeys' welfare and still produces high quality science. NEW METHOD: Here we document refinements in our training methods for acclimation to transport devices (i.e. primate chair or transport box) while working with 46 male rhesus macaques. Our training methods always used positive reinforcement training (PRT). However, PRT was sometimes combined with negative reinforcement training (NRT), but not pole and collar techniques, to successfully transfer each monkey from its home enclosure to its transport device. RESULTS AND COMPARISON WITH EXISTING METHODS: Training monkeys in pairs or groups, and starting their PRT training upon arrival within the unit reduced the days required to acclimate them. While the use of PRT is essential to establish a positive relationship with monkeys, NRT techniques are sometimes necessary, and are most effective when withdrawn immediately once the monkey makes the desired response, to reduce the days of acclimation. Once acclimatised to their chair, monkeys succeeded within 10 days to present their head voluntarily for neck-plating using PRT. Space reducers inside the chairs also facilitated head presentations for some monkeys. CONCLUSIONS: Acclimating (shaping) the monkeys to transport devices can be a stressful experience for monkeys and trainers. The adaptations to our training substantially reduced the days spent on this stage. We view this reduction in days as an effective implementation of the 3Rs (refinement) in monkey neuroscience research.

Patient MW: transient visual hemi-agnosia.

The concept of functional modularity in human visual processing was proposed 25 years ago with the distinction between a ventral pathway for object recognition and a dorsal pathway for action processing. Lesions along these pathways yield selective deficits. A 15-year-old patient (MW) presented with a seizure due to a lesion in the left occipitotemporal cortex. Surgical resection of the lesion was performed with sparing of the classic language areas and visual fields. Postoperatively MW had great difficulty reading and had a specific agnosia for more complex visual stimuli in the right hemifield. No deficit was seen for lower level visual discrimination tasks. Gradual improvement of hemi-agnosia was paralleled by slower reaction times reflecting a speed-accuracy trade-off. Absolute reading speed improved markedly over time, doubling at 6 weeks. MW fully recovered after 18 months. Postoperative functional Magnetic Resonance Imaging (fMRI) illustrated an overlap of the lesion with object and word processing areas. Diffusion Tensor Imaging showed damage to the white matter tracts [inferior fronto-occipital fasciculus and inferior longitudinal fasciculus (ILF)] interconnecting ventral temporal areas. A transient higher order deficit can result from a disruption of the neural network supporting visual word and object processing. Most visual system research has focused on cortical areas, while the underlying subcortical network received much less attention. We believe that white matter tracts, in particular the ILF, play a critical role in object perception by connecting visual areas along the ventral visual stream. Lesions of the ILF should be taken into consideration in agnosia.

Functional MRI in Macaque Monkeys during Task Switching.

Nonhuman primates have proven to be a valuable animal model for exploring neuronal mechanisms of cognitive control. One important aspect of executive control is the ability to switch from one task to another, and task-switching paradigms have often been used in human volunteers to uncover the underlying neuronal processes. To date, however, no study has investigated task-switching paradigms in nonhuman primates during functional magnetic resonance imaging (fMRI). We trained two rhesus macaques to switch between arm movement, eye movement, and passive fixation tasks during fMRI. Similar to results obtained in human volunteers, task switching elicits increased fMRI activations in prefrontal cortex, anterior cingulate cortex, orbitofrontal cortex, and caudate nucleus. Our results indicate that the macaque monkey is a reliable model with which to investigate higher-order cognitive functioning such as task switching. As such, these results can pave the way for a detailed investigation of the neural basis of complex human behavior.SIGNIFICANCE STATEMENT Task switching is an important aspect of cognitive control, and task-switching paradigms have often been used to investigate higher-order executive functioning in human volunteers. We used a task-switching paradigm in the nonhuman primate during fMRI and found increased activation mainly in prefrontal areas (46, 45, frontal eye field, and anterior cingulate), in orbitofrontal area 12, and in the caudate nucleus. These data fit surprisingly well with previous human imaging data, proving that the monkey is an excellent model to study task switching with high spatiotemporal resolution tools that are currently not applicable in humans. As such, our results pave the way for a detailed interrogation of regions performing similar executive functions in humans and monkeys.

Shape responses in a macaque frontal area connected to posterior parietal cortex.

The primate dorsal visual stream processes object shape to guide actions involving an object, but the transmission of shape information beyond posterior parietal cortex remains largely unknown. To clarify the information flow between parietal and frontal cortex, we applied electrical microstimulation during functional Magnetic Resonance Imaging (fMRI) in a shape-selective patch in the posterior part of the Anterior Intraparietal area (pAIP) to chart its connectivity. Subsequently, we recorded single-unit responses to images of objects in the fMRI activation in prefrontal cortex, corresponding to area 45B, elicited by pAIP microstimulation. Neurons in area 45B had properties similar to neurons in pAIP, responding selectively to shape contours and to very small shape fragments measuring less than one deg at exceedingly short latencies. However, contrary to the prevailing view on the hierarchical organization of cortical areas, neurons in area 45B preferred even smaller shape fragments and had smaller receptive fields than neurons in pAIP. These findings provide the first evidence for ultra-fast shape processing in prefrontal cortex, and suggest that the pathway from pAIP to area 45B may not be important for object grasping.

Functional interactions between the macaque dorsal and ventral visual pathways during three-dimensional object vision.

The division of labor between the dorsal and the ventral visual stream in the primate brain has inspired numerous studies on the visual system in humans and in nonhuman primates. However, how and under which circumstances the two visual streams interact is still poorly understood. Here we review evidence from anatomy, modelling, electrophysiology, electrical microstimulation (EM), reversible inactivation and functional imaging in the macaque monkey aimed at clarifying at which levels in the hierarchy of visual areas the two streams interact, and what type of information might be exchanged between the two streams during three-dimensional (3D) object viewing. Neurons in both streams encode 3D structure from binocular disparity, synchronized activity between parietal and inferotemporal areas is present during 3D structure categorization, and clusters of 3D structure-selective neurons in parietal cortex are anatomically connected to ventral stream areas. In addition, caudal intraparietal cortex exerts a causal influence on 3D-structure related activations in more anterior parietal cortex and in inferotemporal cortex. Thus, both anatomical and functional evidence indicates that the dorsal and the ventral visual stream interact during 3D object viewing.

Effective Connectivity Reveals Largely Independent Parallel Networks of Face and Body Patches.

The primate brain processes objects in the ventral visual pathway. One object category, faces, is processed in a hierarchical network of interconnected areas along this pathway. It remains unknown whether such an interconnected network is specific for faces or whether there are similar networks for other object classes. For example, the primate inferotemporal cortex also contains a set of body-selective patches, adjacent to the face-selective patches, but it is not known whether these body-selective patches form a similar discretely connected network or whether cross-talk exists between the face- and body-processing systems. To address these questions, we combined fMRI with electrical microstimulation to determine the effective connectivity of fMRI-defined face and body patches. We found that microstimulation of face patches caused increased fMRI activation throughout the face-processing system; microstimulation of the body patches gave similar results restricted to the body-processing system. Critically, our results revealed largely segregated connectivity patterns for the body and face patches. These results suggest that face and body patches form two interconnected hierarchical networks that are largely separated within the monkey inferotemporal cortex. Only a restricted number of voxels were activated by stimulation of both the body and face patches. The latter regions may be important for the integration of face and body information. Our findings are not only essential to advance our understanding of the neural circuits that enable social cognition, but they also provide further insights into the organizing principles of the inferotemporal cortex.

The effective connectivity of the seizure onset zone and ictal perfusion changes in amygdala kindled rhesus monkeys.

Epileptic seizures are network-level phenomena. Hence, epilepsy may be regarded as a circuit-level disorder that cannot be understood outside this context. Better insight into the effective connectivity of the seizure onset zone and the manner in which seizure activity spreads could lead to specifically-tailored therapies for epilepsy. We applied the electrical amygdala kindling model in two rhesus monkeys until these animals displayed consistent stage IV seizures. At this stage, we investigated the effective connectivity of the amygdala by means of electrical microstimulation during fMRI (EM-fMRI). In addition, we imaged changes in perfusion during a seizure using ictal SPECT perfusion imaging. The spatial overlap between the connectivity network and the ictal perfusion network was assessed both at the regional level, by calculating Dice coefficients using anatomically defined regions of interest, and at the voxel level. The kindled amygdala was extensively connected to bilateral cortical and subcortical structures, which in many cases were connected multisynaptically to the amygdala. At the regional level, the spatial extents of many of these fMRI activations and deactivations corresponded to the respective increases and decreases in perfusion imaged during a stage IV seizure. At the voxel level, however, some regions showed residual seizure-specific activity (not overlapping with the EM-fMRI activations) or fMRI-specific activation (not overlapping with the ictal SPECT activations), indicating that frequently, only a part of a region anatomically connected to the seizure onset zone participated in seizure propagation. Thus, EM-fMRI in the amygdala of electrically-kindled monkeys reveals widespread areas that are often connected multisynaptically to the seizure focus. Seizure activity appears to spread, to a large extent, via these connected areas.

Posterior Parietal Cortex Drives Inferotemporal Activations During Three-Dimensional Object Vision.

The primate visual system consists of a ventral stream, specialized for object recognition, and a dorsal visual stream, which is crucial for spatial vision and actions. However, little is known about the interactions and information flow between these two streams. We investigated these interactions within the network processing three-dimensional (3D) object information, comprising both the dorsal and ventral stream. Reversible inactivation of the macaque caudal intraparietal area (CIP) during functional magnetic resonance imaging (fMRI) reduced fMRI activations in posterior parietal cortex in the dorsal stream and, surprisingly, also in the inferotemporal cortex (ITC) in the ventral visual stream. Moreover, CIP inactivation caused a perceptual deficit in a depth-structure categorization task. CIP-microstimulation during fMRI further suggests that CIP projects via posterior parietal areas to the ITC in the ventral stream. To our knowledge, these results provide the first causal evidence for the flow of visual 3D information from the dorsal stream to the ventral stream, and identify CIP as a key area for depth-structure processing. Thus, combining reversible inactivation and electrical microstimulation during fMRI provides a detailed view of the functional interactions between the two visual processing streams.

Effector specificity in macaque frontal and parietal cortex.

Single neurons in the frontal eye fields (FEFs) and lateral intraparietal area (LIP) of macaques are preferentially activated by saccade- versus reach-related processes. fMRI studies focusing on saccade- and reach-specific activity in human cortex, however, provided conflicting evidence for effector specificity. To gain further insights into effector preferences throughout monkey cortex using the same technique as in humans, we performed a mixed block/event-related fMRI experiment in macaques. Within single fMRI runs, monkeys alternated between a visually guided saccade task, a visually guided arm movement task, and a fixation-only task requiring no saccades or arm movements. The detection of a peripheral pop-out go cue initiating the required operant behavior and the identification of a target among distractors was identical in the arm and saccade tasks. We found saccade-related activity in parietal areas V6, V6A, LIP, and caudal intraparietal area and frontal areas FEF, 45a, 45b, and 46. Areas 45 and FEF even showed markedly decreased fMRI activity during arm movements relative to fixation only. Conversely, medial and anterior intraparietal areas (MIP and AIP), and parietal area PEip; somatosensory areas S1 and S2; and (pre)motor areas F1, F3, F5, and F6 showed increased arm movement-related activity. F1, F5, PEip, and somatosensory cortex also showed deactivations during saccades relative to fixation only. Control experiments showed that such deactivations in both operant-specific functional networks did not depend on training history or rapid task switching requiring active suppression of the unpreferred operant behavior. Therefore, although both tasks required divided attention to detect a pop-out go cue and target, two largely segregated and mainly effector-driven cortical networks were activated.

Effective connectivity of depth-structure-selective patches in the lateral bank of the macaque intraparietal sulcus.

Extrastriate cortical areas are frequently composed of subpopulations of neurons encoding specific features or stimuli, such as color, disparity, or faces, and patches of neurons encoding similar stimulus properties are typically embedded in interconnected networks, such as the attention or face-processing network. The goal of the current study was to examine the effective connectivity of subsectors of neurons in the same cortical area with highly similar neuronal response properties. We first recorded single- and multi-unit activity to identify two neuronal patches in the anterior part of the macaque intraparietal sulcus (IPS) showing the same depth structure selectivity and then employed electrical microstimulation during functional magnetic resonance imaging in these patches to determine the effective connectivity of these patches. The two IPS subsectors we identified-with the same neuronal response properties and in some cases separated by only 3 mm-were effectively connected to remarkably distinct cortical networks in both dorsal and ventral stream in three macaques. Conversely, the differences in effective connectivity could account for the known visual-to-motor gradient within the anterior IPS. These results clarify the role of the anterior IPS as a pivotal brain region where dorsal and ventral visual stream interact during object analysis. Thus, in addition to the anatomical connectivity of cortical areas and the properties of individual neurons in these areas, the effective connectivity provides novel key insights into the widespread functional networks that support behavior.