Modular Organization of Signal Transmission in Primate Somatosensory Cortex.

Axonal patches are known as the major sites of synaptic connections in the cerebral cortex of higher order mammals. However, the functional role of these patches is highly debated. Patches are formed by populations of nearby neurons in a topographic manner and are recognized as the termination fields of long-distance lateral connections within and between cortical areas. In addition, axons form numerous boutons that lie outside the patches, whose function is also unknown. To better understand the functional roles of these two distinct populations of boutons, we compared individual and collective morphological features of axons within and outside the patches of intra-areal, feedforward, and feedback pathways by way of tract tracing in the somatosensory cortex of New World monkeys. We found that, with the exception of tortuosity, which is an invariant property, bouton spacing and axonal convergence properties differ significantly between axons within patch and no-patch domains. Principal component analyses corroborated the clustering of axons according to patch formation without any additional effect by the type of pathway or laminar distribution. Stepwise logistic regression identified convergence and bouton density as the best predictors of patch formation. These findings support that patches are specific sites of axonal convergence that promote the synchronous activity of neuronal populations. On the other hand, no-patch domains could form a neuroanatomical substrate to diversify the responses of cortical neurons.

Network Path Convergence Shapes Low-Level Processing in the Visual Cortex.

Hierarchical counterstream via feedforward and feedback interactions is a major organizing principle of the cerebral cortex. The counterstream, as a topological feature of the network of cortical areas, is captured by the convergence and divergence of paths through directed links. So defined, the convergence degree (CD) reveals the reciprocal nature of forward and backward connections, and also hierarchically relevant integrative properties of areas through their inward and outward connections. We asked if topology shapes large-scale cortical functioning by studying the role of CD in network resilience and Granger causal coupling in a model of hierarchical network dynamics. Our results indicate that topological synchronizability is highly vulnerable to attacking edges based on CD, while global network efficiency depends mostly on edge betweenness, a measure of the connectedness of a link. Furthermore, similar to anatomical hierarchy determined by the laminar distribution of connections, CD highly correlated with causal coupling in feedforward gamma, and feedback alpha-beta band synchronizations in a well-studied subnetwork, including low-level visual cortical areas. In contrast, causal coupling did not correlate with edge betweenness. Considering the entire network, the CD-based hierarchy correlated well with both the anatomical and functional hierarchy for low-level areas that are far apart in the hierarchy. Conversely, in a large part of the anatomical network where hierarchical distances are small between the areas, the correlations were not significant. These findings suggest that CD-based and functional hierarchies are interrelated in low-level processing in the visual cortex. Our results are consistent with the idea that the interplay of multiple hierarchical features forms the basis of flexible functional cortical interactions.

Synaptic organization of cortico-cortical communication in primates.

In cortical circuitry, synaptic communication across areas is based on two types of axon terminals, small and large, with modulatory and driving roles, respectively. In contrast, it is not known whether similar synaptic specializations exist for intra-areal projections. Using anterograde tracing and three-dimensional reconstruction by electron microscopy (3D-EM), we asked whether large boutons form synapses in the circuit of somatosensory cortical areas 3b and 1. In contrast to observations in macaque visual cortex, light microscopy showed both small and large boutons not only in inter-areal pathways, but also in long-distance intrinsic connections. 3D-EM showed that correlation of surface and volume provides a powerful tool for classifying cortical endings. Principal component analysis supported this observation and highlighted the significance of the size of mitochondria as a distinguishing feature of bouton type. The larger mitochondrion and higher degree of perforated postsynaptic density associated with large rather than to small boutons support the driver-like function of large boutons. In contrast to bouton size and complexity, the size of the postsynaptic density appeared invariant across the bouton types. Comparative studies in human supported that size is a major distinguishing factor of bouton type in the cerebral cortex. In conclusion, the driver-like function of the large endings could facilitate fast dissemination of tactile information within the intrinsic and inter-areal circuitry of areas 3b and 1.

Corrigendum to "Structural correlates of modular organization of activity propagation in the primate somatosensory cortex" [IBROR 6S (2019) S540].

[This corrects the article DOI: 10.1016/j.ibror.2019.07.1681.].

[Neuronal connections within the hand representation in areas 3b and 1 of the somatosensory cortex in primates]. / Neuronalis összeköttetések a szomatoszenzoros kérgi área 3b és área 1 kézreprezentációs területén foemlosökben.

INTRODUCTION: The close functional relationship between areas 3b and 1 of the somatosensory cortex is based on their reciprocal connections indicating that tactile sensation depends on the interaction of these two areas. AIM: The aim of the authors was to explore this neuronal circuit at the level of the distal finger pad representation. METHOD: The study was made by bidirectional tract tracing aided by neurophysiological mapping in squirrel monkeys (Saimiri sciureus). RESULTS: Inter-areal connections between the two areas preferred the homologues representations. However, intra-areal connections were formed between the neighboring finger pad representations supporting the physiological observations. Interestingly, the size of the local input area of the injected cortical micro-region, which differed in the two areas, represented the same skin area. CONCLUSIONS: The authors propose that intra-areal connections are important in integrating information across fingers, while inter-areal connections are important in maintaining input localization during hand movement. Orv. Hetil., 2016, 157(33), 1320-1325.

Rediscovering TNAP in the Brain: A Major Role in Regulating the Function and Development of the Cerebral Cortex.

The presence of alkaline phosphatase (AP) activity in the neural tissue has been described decades ago. However, only recent studies clarified the isotype, regional distribution and subcellular localization of the AP expressed in the cerebral cortex of diverse mammalian species including the human. In the primate brain the discovery that the bone AP isotype (TNAP) is expressed provided the opportunity of a deeper understanding of the role of this enzyme in neuronal functions based on the knowledge acquired by studying the role of the enzyme in hypophosphatasia, mostly in bone mineralization. TNAP exhibits widespread substrate specificity and, in the brain, it is potentially involved in the regulation of molecules which play fundamental roles in signal transmission and development. In light of these observations, the localization of TNAP in the human cerebral cortex is of high significance when considering that epilepsy is often diagnosed in hypophosphatasia. Here we overview our results on the identification of TNAP in the primate cerebral cortex: TNAP exhibits a noticeably high activity in the synapses and nodes of Ranvier, is specifically present in layer 4 of the sensory cortices and additionally in layer 5 of prefrontal, temporal and other associational areas in human. Our studies also indicate that bone AP activity depends on the level of sensory input and that its developmental time-course exhibits characteristic regional differences. The relevance of our findings regarding human cortical physiology and brain disorders are discussed.

Signal Transduction Pathways of TNAP: Molecular Network Analyses.

Despite the growing body of evidence pointing on the involvement of tissue non-specific alkaline phosphatase (TNAP) in brain function and diseases like epilepsy and Alzheimer's disease, our understanding about the role of TNAP in the regulation of neurotransmission is severely limited. The aim of our study was to integrate the fragmented knowledge into a comprehensive view regarding neuronal functions of TNAP using objective tools. As a model we used the signal transduction molecular network of a pyramidal neuron after complementing with TNAP related data and performed the analysis using graph theoretic tools. The analyses show that TNAP is in the crossroad of numerous pathways and therefore is one of the key players of the neuronal signal transduction network. Through many of its connections, most notably with molecules of the purinergic system, TNAP serves as a controller by funnelling signal flow towards a subset of molecules. TNAP also appears as the source of signal to be spread via interactions with molecules involved among others in neurodegeneration. Cluster analyses identified TNAP as part of the second messenger signalling cascade. However, TNAP also forms connections with other functional groups involved in neuronal signal transduction. The results indicate the distinct ways of involvement of TNAP in multiple neuronal functions and diseases.

Stratified organization and disorganization of inner plexiform layer revealed by TNAP activity in healthy and diabetic rat retina.

Tissue non-specific alkaline phosphatase (TNAP), an abundant ectophosphatase, is present in various organs including the brain and retina of several vertebrate species. Evidence is emerging that TNAP influences neural functions in multiple ways. In rat, strong TNAP activity has been found in retinal vessels, photoreceptors, and both synaptic layers. In the present study, we identified eleven strata of the inner plexiform layer (IPL) by using TNAP histochemistry alone. The TNAP strata corresponded exactly to the strata seen after combined immunohistochemistry with four canonical IPL markers (TH-ChAT-CR-PKCα). Therefore, as described in other mammalian species, our data support the existence of multiple morphologically and functionally discernible IPL strata in rats. Remarkably, the stratification pattern of the IPL was severely disrupted in a diabetic rat model, even before changes in the canonical IPL markers were detectable. These findings indicate that TNAP histochemistry offers a more straightforward, but also more sensitive, method for investigating retinal strata and their diabetes-induced degeneration.

TNAP activity is localized at critical sites of retinal neurotransmission across various vertebrate species.

Evidence is emerging with regard to the role of tissue non-specific alkaline phosphatase (TNAP) in neural functions. As an ectophosphatase, this enzyme might influence neural activity and synaptic transmission in diverse ways. The localization of the enzyme in known neural circuits, such as the retina, might significantly advance an understanding of its role in normal and pathological functioning. However, the presence of TNAP in the retina is scarcely investigated. Our multispecies comparative study (zebrafish, cichlid, frog, chicken, mouse, rat, golden hamster, guinea pig, rabbit, sheep, cat, dog, ferret, squirrel monkey, human) using enzyme histochemistry and Western blots has shown the presence of TNAP activity in the retina of several mammalian species, including humans. Although the TNAP activity pattern varies across species, we have observed the following trends: (1) in all investigated species (except golden hamster), retinal vessels display TNAP activity; (2) TNAP activity consistently occurs in the photoreceptor layer; (3) in majority of the investigated species, marked TNAP activity is present in the outer and inner plexiform layers. In zebrafish, frog, chicken, guinea pig, and rat, TNAP histochemistry has revealed several sublayers of the inner plexiform layer. Frog, golden hamster, guinea pig, mouse, and human retinas possess a subpopulation of amacrine cells positively staining for TNAP activity. The expression of TNAP in critical sites of retinal signal transmission across a wide range of species suggests its fundamental, evolutionally conserved role in vision.

Connectivity of somatosensory cortical area 1 forms an anatomical substrate for the emergence of multifinger receptive fields and complex feature selectivity in the squirrel monkey (Saimiri sciureus).

Converging evidence shows that interaction of digit-specific input, which is required to form global tactile percepts, begins as early as area 3b in the primary somatosensory cortex with the involvement of intrinsic lateral connections. How tactile processing is further elaborated in area 1, the next stage of the somatosensory cortical hierarchy, is less understood. This question was investigated by studying the tangential distribution of intrinsic and interareal connections of finger representations of area 1. Retrogradely labeled cell densities and anterogradely labeled fibers and terminal patches were plotted and quantified with respect to the hand representation by combining tract tracing with electrophysiological mapping and intrinsic signal optical imaging in somatosensory areas. Intrinsic connections of distal finger pad representations of area 1 spanned the representation of multiple digits indicating strong cross-digit connectivity. Area 1 distal finger pad regions also established high-density connections with homotopic regions of areas 3b and 2. Although similar to area 3b, connections of area 1 distributed more widely and covered a larger somatotopic representation including more proximal parts of the finger representations. The lateral connectivity pattern of area 1 is a suitable anatomical substrate of the emergence of multifinger receptive fields, complex feature selectivity, and invariant stimulus properties of the neurons.

The relationship of anatomical and functional connectivity to resting-state connectivity in primate somatosensory cortex.

Studies of resting-state activity in the brain have provoked critical questions about the brain's functional organization, but the biological basis of this activity is not clear. Specifically, the relationships between interregional correlations in resting-state measures of activity, neuronal functional connectivity and anatomical connectivity are much debated. To investigate these relationships, we have examined both anatomical and steady-state functional connectivity within the hand representation of primary somatosensory cortex (areas 3b and 1) in anesthetized squirrel monkeys. The comparison of three data sets (fMRI, electrophysiological, and anatomical) indicate two primary axes of information flow within the SI: prominent interdigit interactions within area 3b and predominantly homotopic interactions between area 3b and area 1. These data support a strikingly close relationship between baseline functional connectivity and anatomical connections. This study extends findings derived from large-scale cortical networks to the realm of local millimeter-scale networks.

Intrinsic horizontal connections process global tactile features in the primary somatosensory cortex: neuroanatomical evidence.

To understand manual tactile functions in primates, it is essential to explore the interactions between the finger pad representations in somatosensory cortex. To this end, we used optical imaging and electrophysiological mapping to guide neuroanatomical tracer injections into distal digit tip representations of Brodmann area 3b in the squirrel monkey. Retrogradely labeled cell densities and anterogradely labeled fibers and terminal patches in somatosensory areas were plotted and quantified with respect to tangential distribution. Within area 3b, reciprocal patchy distribution of anterograde and retrograde labeling spanned the representation of the distal pad of multiple digits, indicating strong cross-digit connectivity. Inter-areal connections revealed bundles of long-range fibers projecting anteroposteriorly, connecting area 3b with clusters of labeled neurons and terminal axon arborizations in area 1. Inter-areal linkage appeared to be largely confined to the representation of the injected finger. These findings provide the neuroanatomical basis for the interaction between distal finger pad representations observed by recent electrophysiological studies. We propose that intra-areal connectivity may be heavily involved in interdigit integration such as shape discrimination, whereas long-range inter-areal connections may subserve active touch in a digit-specific manner.

Neurochemical mapping of the human hippocampus reveals perisynaptic matrix around functional synapses in Alzheimer's disease.

Perineuronal matrix is an extracellular protein scaffold to shape neuronal responsiveness and survival. Whilst perineuronal nets engulf the somatodendritic axis of neurons, axonal coats are focal extracellular protein aggregates surrounding individual synapses. Here, we addressed the chemical identity and subcellular localization of both perineuronal and perisynaptic matrices in the human hippocampus, whose neuronal circuitry is progressively compromised in Alzheimer's disease. We hypothesized that (1) the cellular expression sites of chondroitin sulphate proteoglycan-containing extracellular matrix associate with specific neuronal identities, reflecting network dynamics, and (2) the regional distribution and molecular composition of axonal coats must withstand Alzheimer's disease-related modifications to protect functional synapses. We show by epitope-specific antibodies that the perineuronal protomap of the human hippocampus is distinct from other mammals since pyramidal cells but not calretinin(+) and calbindin(+) interneurons, neurochemically classified as novel neuronal subtypes, lack perineuronal nets. We find that cartilage link protein-1 and brevican-containing matrices form isolated perisynaptic coats, engulfing both inhibitory and excitatory terminals in the dentate gyrus and entorhinal cortex. Ultrastructural analysis revealed that presynaptic neurons contribute components of perisynaptic coats via axonal transport. We demonstrate, by combining biochemical profiling and neuroanatomy in Alzheimer's patients and transgenic (APdE9) mice, the preserved turnover and distribution of axonal coats around functional synapses along dendrite segments containing hyperphosphorylated tau and in amyloid-ß-laden hippocampal microdomains. We conclude that the presynapse-driven formation of axonal coats is a candidate mechanism to maintain synapse integrity under neurodegenerative conditions.

Ablation of TNAP function compromises myelination and synaptogenesis in the mouse brain.

Mutations in the tissue-nonspecific alkaline phosphatase (TNAP) gene can result in skeletal and dental hypomineralization and severe neurological symptoms. TNAP is expressed in the synaptic cleft and the node of Ranvier in normal adults. Using TNAP knockout (KO) mice (Akp2(-/-)), we studied synaptogenesis and myelination with light- and electron microscopy during the early postnatal days. Ablation of TNAP function resulted in a significant decrease of the white matter of the spinal cord accompanied by ultrastructural evidence of cellular degradation around the paranodal regions and a decreased ratio and diameter of the myelinated axons. In the cerebral cortex, myelinated axons, while present in wild-type, were absent in the Akp2( -/- ) mice and these animals also displayed a significantly increased proportion of immature cortical synapses. The results suggest that TNAP deficiency could contribute to neurological symptoms related to myelin abnormalities and synaptic dysfunction, among which epilepsy, consistently present in the Akp2(-/-) mice and observed in severe cases of hypophosphatasia.

Distribution and classification of aggrecan-based extracellular matrix in the thalamus of the rat.

Extracellular matrix molecules take part in functional isolation and stabilization of neuronal compartments but form a vivid interface between neuronal elements at the same time. Previous studies have shown that the accumulation of extracellular matrix, especially its typical phenotypic form, termed perineuronal nets, correlates not only with the functional properties of the single neuron but also with the functional properties of the whole brain area. In contrast to recent advances in investigating neocortex, the present study mapped the occurrence and phenotypic appearance of aggrecan-based matrix accumulation throughout the rat thalamus. Results showed that divisions of thalamus that relay information to cortical fields known rather for their plastic properties exibit a poor matrix immunoreactivity, whereas matrix accumulation is more enhanced in nuclei connected to primary cortical regions. In addition to perineuronal nets, extracellular matrix condensed in another peculiar form, in 2-5-µm, large, round or oval structures, as described by Brückner et al. ([ 2008] Neuroscience 151:489-504) as axonal coats (ACs). Multiple labelling experiments showed that specific excitatory afferents were not ensheathed with these structures. At the same time, inhibitory endings were occasionally enwrapped in ACs. Electron microscopic analysis showed that aggrecan-immunoreactive profiles were present mostly around inhibitory terminals but also in all neuronal compartments. We suggest that aggrecan-based extracellular matrix is formed by both pre- and postsynaptic elements and is preferably associated with inhibitory terminals in the extracellular space.

Convergence and divergence are mostly reciprocated properties of the connections in the network of cortical areas.

Cognition is based on the integrated functioning of hierarchically organized cortical processing streams in a manner yet to be clarified. Because integration fundamentally depends on convergence and the complementary notion of divergence of the neuronal connections, we analysed integration by measuring the degree of convergence/divergence through the connections in the network of cortical areas. By introducing a new index, we explored the complementary convergent and divergent nature of connectional reciprocity and delineated the backward and forward cortical sub-networks for the first time. Integrative properties of the areas defined by the degree of convergence/divergence through their afferents and efferents exhibited distinctive characteristics at different levels of the cortical hierarchy. Areas previously identified as hubs exhibit information bottleneck properties. Cortical networks largely deviate from random graphs where convergence and divergence are balanced at low reciprocity level. In the cortex, which is dominated by reciprocal connections, balance appears only by further increasing the number of reciprocal connections. The results point to the decisive role of the optimal number and placement of reciprocal connections in large-scale cortical integration. Our findings also facilitate understanding of the functional interactions between the cortical areas and the information flow or its equivalents in highly recurrent natural and artificial networks.

Ultrastructural localization of calcyon in the primate cortico-basal ganglia-thalamocortical loop.

Recent observations suggest that calcyon, a novel single transmembrane protein implicated in schizophrenia and attention-deficit/hyperactivity disorder, regulates clathrin-mediated endocytosis in brain. To explore the role of calcyon in neurotransmission, we investigated its distribution in the neuropil of the primate prefrontal cortex (PFC), striatum (STR) and mediodorsal thalamic nucleus (MD), three brain regions implicated in these neuropsychiatric disorders. Calcyonimmunoreactivity revealed by immunoperoxidase technique, was localized in both pre- and postsynaptic structures including axons, spines and dendrites, as well as myelinated fibers and astroglial processes in all the three brain regions. The morphological diversity of immunopositive boutons suggest that in addition to glutamatergic, calcyon could regulate GABAergic as well as monoaminergic neurotransmission. Consistent with the role of calcyon in endocytosis, calcyon-immunoreactivity was rarely found at the synaptic membrane specializations proper, although it was present in distal compartments of neuronal processes establishing synapses. Given the widespread upregulation of calcyon in schizophrenic brain, these findings underscore a potential association with deficits in a range of neurotransmitter systems in the cortico-basal ganglia-thalamic loop.

Fuzzy communities and the concept of bridgeness in complex networks.

We consider the problem of fuzzy community detection in networks, which complements and expands the concept of overlapping community structure. Our approach allows each vertex of the graph to belong to multiple communities at the same time, determined by exact numerical membership degrees, even in the presence of uncertainty in the data being analyzed. We create an algorithm for determining the optimal membership degrees with respect to a given goal function. Based on the membership degrees, we introduce a measure that is able to identify outlier vertices that do not belong to any of the communities, bridge vertices that have significant membership in more than one single community, and regular vertices that fundamentally restrict their interactions within their own community, while also being able to quantify the centrality of a vertex with respect to its dominant community. The method can also be used for prediction in case of uncertainty in the data set analyzed. The number of communities can be given in advance, or determined by the algorithm itself, using a fuzzified variant of the modularity function. The technique is able to discover the fuzzy community structure of different real world networks including, but not limited to, social networks, scientific collaboration networks, and cortical networks, with high confidence.

Prediction of the main cortical areas and connections involved in the tactile function of the visual cortex by network analysis.

We explored the cortical pathways from the primary somatosensory cortex to the primary visual cortex (V1) by analysing connectional data in the macaque monkey using graph-theoretical tools. Cluster analysis revealed the close relationship of the dorsal visual stream and the sensorimotor cortex. It was shown that prefrontal area 46 and parietal areas VIP and 7a occupy a central position between the different clusters in the visuo-tactile network. Among these structures all the shortest paths from primary somatosensory cortex (3a, 1 and 2) to V1 pass through VIP and then reach V1 via MT, V3 and PO. Comparison of the input and output fields suggested a larger specificity for the 3a/1-VIP-MT/V3-V1 pathways among the alternative routes. A reinforcement learning algorithm was used to evaluate the importance of the aforementioned pathways. The results suggest a higher role for V3 in relaying more direct sensorimotor information to V1. Analysing cliques, which identify areas with the strongest coupling in the network, supported the role of VIP, MT and V3 in visuo-tactile integration. These findings indicate that areas 3a, 1, VIP, MT and V3 play a major role in shaping the tactile information reaching V1 in both sighted and blind subjects. Our observations greatly support the findings of the experimental studies and provide a deeper insight into the network architecture underlying visuo-tactile integration in the primate cerebral cortex.

Calcyon, a novel partner of clathrin light chain, stimulates clathrin-mediated endocytosis.

In the central nervous system, clathrin-mediated endocytosis is crucial for efficient synaptic transmission. Clathrin-coated vesicle assembly and disassembly is regulated by some 30 adaptor and accessory proteins, most of which interact with clathrin heavy chain. Using the calcyon cytosolic domain as bait, we isolated clathrin light chain in a yeast two-hybrid screen. The interaction domain was mapped to the heavy chain binding domain and C-terminal regions of light chain. Further, the addition of the calcyon C terminus stimulated clathrin self-assembly in a dose-dependent fashion. Calcyon, which is a single transmembrane protein predominantly expressed in brain, localized to vesicular compartments within pre- and postsynaptic structures. There was a high degree of overlap in the distribution of LC and calcyon in neuronal dendrites, spines, and cell bodies. Co-immunoprecipitation studies further suggested an association of calcyon with the clathrin-mediated endocytic machinery. Compared with controls, HEK293 cells overexpressing calcyon exhibited significantly enhanced transferrin uptake but equivalent levels of recycling. Conversely, transferrin uptake was largely abolished in neocortical neurons obtained from mice homozygous for a calcyon null allele, whereas recycling proceeded at wild type levels. Collectively, these data indicate a role for calcyon in clathrin-mediated endocytosis in brain.