Polysynaptic subcircuits in the neocortex: spatial and temporal diversity

Inhibitory pathways in the neocortex display a variety of temporal and spatial patterns, maintaining a dynamic balance with excitatory synaptic activity. Recent studies have revealed prevalent polysynaptic subcircuits within the neocortical microcircuitry. These subcircuits involve excitatory and inhibitory connections that are activated by neurons both in supragranular and infragranular cortical layers and mediated by different mechanisms. Interestingly, in these subcircuits inhibition is induced by discharge of pyramidal cells, and excitation is caused by specific types of GABAergic interneurons. The different polysynaptic subcircuits are discussed with respect to their spatial and temporal properties and their possible functional role in cortical processing.     

Light microscopy of the medial wall of the cerebral cortex of the lizard Psammodromus algirus

The telencephalic medial wall of the lizard Psammodromus algirus was studied using Golgi and conventional light microscopic techniques. The area is formed by two different cytological fields—medial cortex and dorsomedial cortex. These two cortices possess three layers dorsoventrally: a superficial plexiform layer, a cellular layer, and a deep plexiform layer. The alveus, a deep fiber system, runs adjacent to the ependyma. Four classes of neurons are found in the cellular layer of the medial cortex on the basis of soma shape, dendritic pattern, and position in the layer: horizontal, double pyramidal, and candelabra cells. Solitary cells are present in the superficial and deep plexiform layers of the medial cortex. Those of the superficial plexiform layer are stellate cells. Horizontal and vertical cells are found in the deep plexiform layer. Double pyramidal cells are the most frequently impregnated in the cellular layer of the dorsomedial cortex. In addition, candelabra cells are present at the lateral end of the layer. Two cell types are found in the deep plexiform layer of the dorsomedial cortex: solitary pyramidal cells and, among the fibers of the alveus, horizontal cells. Ependymal tanycytes line the ventricular surface, and protoplasmic astrocytes are found in the plexiform layers of both medial and dorsomedial cortices.     

Brief Bursts Self-Inhibit and Correlate the Pyramidal Network

Inhibitory pathways are an essential component in the function of the neocortical microcircuitry. Despite the relatively small fraction of inhibitory neurons in the neocortex, these neurons are strongly activated due to their high connectivity rate and the intricate manner in which they interconnect with pyramidal cells (PCs). One prominent pathway is the frequency-dependent disynaptic inhibition (FDDI) formed between layer 5 PCs and mediated by Martinotti cells (MCs). Here, we show that simultaneous short bursts in four PCs are sufficient to exert FDDI in all neighboring PCs within the dimensions of a cortical column. This powerful inhibition is mediated by few interneurons, leading to strongly correlated membrane fluctuations and synchronous spiking between PCs simultaneously receiving FDDI. Somatic integration of such inhibition is independent and electrically isolated from monosynaptic excitation formed between the same PCs. FDDI is strongly shaped by I(h) in PC dendrites, which determines the effective integration time window for inhibitory and excitatory inputs. We propose a key disynaptic mechanism by which brief bursts generated by a few PCs can synchronize the activity in the pyramidal network.     

Laminar and Dorsoventral Molecular Organization of the Medial Entorhinal Cortex Revealed by Large-scale Anatomical Analysis of Gene Expression

Neural circuits in the medial entorhinal cortex (MEC) encode an animal’s position and orientation in space. Within the MEC spatial representations, including grid and directional firing fields, have a laminar and dorsoventral organization that corresponds to a similar topography of neuronal connectivity and cellular properties. Yet, in part due to the challenges of integrating anatomical data at the resolution of cortical layers and borders, we know little about the molecular components underlying this organization. To address this we develop a new computational pipeline for high-throughput analysis and comparison of in situ hybridization (ISH) images at laminar resolution. We apply this pipeline to ISH data for over 16,000 genes in the Allen Brain Atlas and validate our analysis with RNA sequencing of MEC tissue from adult mice. We find that differential gene expression delineates the borders of the MEC with neighboring brain structures and reveals its laminar and dorsoventral organization. We propose a new molecular basis for distinguishing the deep layers of the MEC and show that their similarity to corresponding layers of neocortex is greater than that of superficial layers. Our analysis identifies ion channel-, cell adhesion- and synapse-related genes as candidates for functional differentiation of MEC layers and for encoding of spatial information at different scales along the dorsoventral axis of the MEC. We also reveal laminar organization of genes related to disease pathology and suggest that a high metabolic demand predisposes layer II to neurodegenerative pathology. In principle, our computational pipeline can be applied to high-throughput analysis of many forms of neuroanatomical data. Our results support the hypothesis that differences in gene expression contribute to functional specialization of superficial layers of the MEC and dorsoventral organization of the scale of spatial representations.     

Sonic hedgehog expression in corticofugal projection neurons directs cortical microcircuit formation

The precise connectivity of inputs and outputs is critical for cerebral cortex function; however, the cellular mechanisms that establish these connections are poorly understood. Here, we show that the secreted molecule Sonic Hedgehog (Shh) is involved in synapse formation of a specific cortical circuit. Shh is expressed in layer V corticofugal projection neurons and the Shh receptor, Brother of CDO (Boc), is expressed in local and callosal projection neurons of layer II/III that synapse onto the subcortical projection neurons. Layer V neurons of mice lacking functional Shh exhibit decreased synapses. Conversely, the loss of functional Boc leads to a reduction in the strength of synaptic connections onto layer Vb, but not layer II/III, pyramidal neurons. These results demonstrate that Shh is expressed in postsynaptic target cells while Boc is expressed in a complementary population of presynaptic input neurons, and they function to guide the formation of cortical microcircuitry. VIDEO ABSTRACT:     

The time window for generation of dendritic spikes by coincidence of action potentials and EPSPs is layer specific in somatosensory cortex

The precise timing of events in the brain has consequences for intracellular processes, synaptic plasticity, integration and network behaviour. Pyramidal neurons, the most widespread excitatory neuron of the neocortex have multiple spike initiation zones, which interact via dendritic and somatic spikes actively propagating in all directions within the dendritic tree. For these neurons, therefore, both the location and timing of synaptic inputs are critical. The time window for which the backpropagating action potential can influence dendritic spike generation has been extensively studied in layer 5 neocortical pyramidal neurons of rat somatosensory cortex. Here, we re-examine this coincidence detection window for pyramidal cell types across the rat somatosensory cortex in layers 2/3, 5 and 6. We find that the time-window for optimal interaction is widest and shifted in layer 5 pyramidal neurons relative to cells in layers 6 and 2/3. Inputs arriving at the same time and locations will therefore differentially affect spike-timing dependent processes in the different classes of pyramidal neurons.     

Experience-Dependent Rewiring of Specific Inhibitory Connections in Adult Neocortex

Although neocortical connectivity is remarkably stereotyped, the abundance of some wiring motifs varies greatly between cortical areas. To examine if regional wiring differences represent functional adaptations, we have used optogenetic raster stimulation to map the laminar distribution of GABAergic interneurons providing inhibition to pyramidal cells in layer 2/3 (L2/3) of adult mouse barrel cortex during sensory deprivation and recovery. Whisker trimming caused large, motif-specific changes in inhibitory synaptic connectivity: ascending inhibition from deep layers 4 and 5 was attenuated to 20%–45% of baseline, whereas inhibition from superficial layers remained stable (L2/3) or increased moderately (L1). The principal mechanism of deprivation-induced plasticity was motif-specific changes in inhibitory-to-excitatory connection probabilities; the strengths of extant connections were left unaltered. Whisker regrowth restored the original balance of inhibition from deep and superficial layers. Targeted, reversible modifications of specific inhibitory wiring motifs thus contribute to the adaptive remodeling of cortical circuits.     

Changes of Dendritic Spine Density and Morphology in the Superficial Layers of the Medial Entorhinal Cortex Induced by Extremely Low-Frequency Magnetic Field Exposure

In the present study, we investigated the effects of chronic exposure (14 and 28 days) to a 0.5 mT 50 Hz extremely low-frequency magnetic field (ELM) on the dendritic spine density and shape in the superficial layers of the medial entorhinal cortex (MEC). We performed Golgi staining to reveal the dendritic spines of the principal neurons in rats. The results showed that ELM exposure induced a decrease in the spine density in the dendrites of stellate neurons and the basal dendrites of pyramidal neurons at both 14 days and 28 days, which was largely due to the loss of the thin and branched spines. The alteration in the density of mushroom and stubby spines post ELM exposure was cell-type specific. For the stellate neurons, ELM exposure slightly increased the density of stubby spines at 28 days, while it did not affect the density of mushroom spines at the same time. In the basal dendrites of pyramidal neurons, we observed a significant decrease in the mushroom spine density only at the later time point post ELM exposure, while the stubby spine density was reduced at 14 days and partially restored at 28 days post ELM exposure. ELM exposure-induced reduction in the spine density in the apical dendrites of pyramidal neurons was only observed at 28 days, reflecting the distinct vulnerability of spines in the apical and basal dendrites. Considering the changes in spine number and shape are involved in synaptic plasticity and the MEC is a part of neural network that is closely related to learning and memory, these findings may be helpful for explaining the ELM exposure-induced impairment in cognitive functions.     

The structure of the first auditory cortex (A I) in the cat. I.--Light microscopic observations on its organization

The cyto- and mieloarchitecture of the first auditory cortex (A I) was studied in the cat. The cortical layers II, III and IV are very densely populated by relatively uniform, round or stellate cells with 20 to 30 micro perikaryal diameter. The separation between these three layers, which is not possible in Nissl stained sections, becomes visible in 1 to 3 micro thick sections of plastic embedded material. nerve cells in layer II are randomly disposed, whilst they form in laver III loose rounded cellular groups, and in layer IV vertical cylinders which have 50 to 60 micro in outside diameter and a cell poor centre. These cylinders are best visible in 100 micro thick Nissl preparations, cut parallel to the pial surface. The cylinders may extend into layer V, which is comparatively cell poor. The VIth layer contains numerous round, stellate or fusiform cells with 20 to 30 micro in diameter. The IIIrd and Vth layers have few pyramidal perikarya which are small. Large or giant pyramidal cells are not found in A I. The overall thickness of the cortex in the convexity of A I is 2,000 micro, measured in sections of plastic blocks. The thickness of the 6 layers is 200 to 250 micro for layer I; 300 micro for layer II; 300 micro for layer III; 300 to 400, for layer IV; 350 micro for layer V; and 400 micro for layer VI. In preparations stained for myelin sheats A I is characterized by the presence of a very dense plexus of fibres running in all directions in the IVth, Vth anti VIth layers. These plexus obscurs the radiations of Meynert, giving a characteristic appearance to A I, since these radiations are prominent in the neighbouring cortical areas. In preliminary studies of Golgi rapid preparations of A I the cell types commonly present in others cortical areas were found. Pyramidal cells have small perikarya, and very long (600 micro) horizontal basal dendrites. Modified pyramidal cells (star pyramids) are the main cellular element in layer II and constitute one of the main sources of efferent fibres of A I. Several types of stellate cells were found, including a particular cell type, found very often in the IVth layer, with a very long horizontal axon. The specific thalamic afferents were identified as fibres with 5 or 8 micro in diameter, which run obliquely and sinuously through the VIth and Vth layers of A I. These fibres give off many branches with 1 to 2 micro in diameter, which pass to the IVth layer where they give off very thin sinuous branches, ending in small terminal knobs. The ramification of one of these fibres may spread horizontally over 800 micros, at the level of the IVth layer.     

Pyramidal Cells Make Specific Connections onto Smooth (GABAergic) Neurons in Mouse Visual Cortex

One of the hallmarks of neocortical circuits is the predominance of recurrent excitation between pyramidal neurons, which is balanced by recurrent inhibition from smooth GABAergic neurons. It has been previously described that in layer 2/3 of primary visual cortex (V1) of cat and monkey, pyramidal cells filled with horseradish peroxidase connect approximately in proportion to the spiny (excitatory, 95% and 81%, respectively) and smooth (GABAergic, 5% and 19%, respectively) dendrites found in the neuropil. By contrast, a recent ultrastructural study of V1 in a single mouse found that smooth neurons formed 51% of the targets of the superficial layer pyramidal cells. This suggests that either the neuropil of this particular mouse V1 had a dramatically different composition to that of V1 in cat and monkey, or that smooth neurons were specifically targeted by the pyramidal cells in that mouse. We tested these hypotheses by examining similar cells filled with biocytin in a sample of five mice. We found that the average composition of the neuropil in V1 of these mice was similar to that described for cat and monkey V1, but that the superficial layer pyramidal cells do form proportionately more synapses with smooth dendrites than the equivalent neurons in cat or monkey. These distributions may underlie the distinct differences in functional architecture of V1 between rodent and higher mammals.     

Modulatory effects of parallel fiber and molecular layer interneuron synaptic activity on purkinje cell responses to ascending segment input: a modeling study

Based on anatomical, physiological, and model-based studies, it has been proposed that synapses associated with the ascending segment of granule cell axons provide the principle excitatory drive on Purkinje cells which is then modulated by the more numerous parallel fiber synapses. In this study we have evaluated this idea using a detailed compartmental model of a cerebellar Purkinje cell by providing identical ascending segment synaptic inputs during different levels of random parallel fiber and molecular interneuron input. Results suggest that background inputs from parallel fibers and molecular layer interneurons can have a substantial effect on the response of Purkinje cells to ascending segment inputs. Interestingly, these effects are not reflected in the average firing rate of the Purkinje cell and are thus entirely dendritic in effect. These results are considered in the context of the known segregated spatial distribution of the parallel fibers and ascending segment synapses and a new hypothesis concerning the functional organization of cerebellar cortical circuitry.     

Neurons and their interconnections in the frontal cortex of monkeys

It was shown by the Golgi and Golgi-Kopsch method that pyramidal cells of layers II–IV in the frontal cortex of the monkeyMacaca rhesus have numeruous, mainly recurrent axon collaterals by means of which they form vertical connections. Pyramidal cells with ascending axons are found. Axons of stellate basket neurons unite pyramidal cells in both horizontal (modules) and vertical (micromodules) directions; depending on the direction of the axon collaterals, two groups of stellate neurons can be distinguished. Groups of 14 to 16 pyramidal cells whose apical dendrites are connected into bundles were found. Axons of pyramidal cells in layers II–IV descend in the composition of the pyramidal tract and give off collaterals which run toward the bodies and dendrites of neighboring pyramidal cells, united into the same group, forming terminal and en passant junctions. Besides bundles, special kinds of "local" cell groups with U-shaped axons are found.A. A. Zhdanov Leningrad State University. Translated from Neirofiziologiya, Vol. 15, No. 2, pp. 115–120, March–April, 1983.     

Expression of nerve growth factor and its receptors in the somatosensory-motor cortex of Macaca nemestrina

The present study was designed to examine the nerve growth factor (NGF) system (ligand and receptor-expressing neurons) in the somatosensory (areas 1, 3a, and 3b) and motor (area 4) cortices of the mature macaque. Light and electron microscope immunohistochemistry was used to assess the distribution and identity of NGF-, p75-, and trk-expressing elements. In each cortical area examined, NGF-positive neuronal somata were distributed through all laminae; most immunolabeled neurons were in layers II, III, and V. Based upon light microscope criteria (e.g., the morphology of proximal dendrites), both pyramidal and stellate neurons expressed NGF. Of the identifiable NGF- immunoreactive cells, 92% were pyramidal neurons and the remainder was stellate neurons. The electron microscope study showed that most (88%) NGF-positive somata formed symmetric synapses, whereas the others formed both symmetric and asymmetric synapses. As the somata of pyramidal neurons form only symmetric synapses and those of inhibitory stellate neurons form both symmetric and asymmetric somatic synapses, the ultrastructural data support the light microscopic analyses. In contrast, neurotrophin receptors, p75 and trk, were expressed chiefly by the cell bodies of layer V pyramidal neurons and the supragranular neuropil. At the ultrastructural level, receptor-positive profiles were post-synaptic elements (e.g., dendritic shafts and spines) and the concentration of immunoreactivity was greatest in the vicinity of post-synaptic densities. Thus, NGF regulatory systems parallel excitatory and inhibitory neurotransmitter systems. Cortex contains the morphological framework by which pyramidal and/or inhibitory stellate neurons can affect the activity of post-synaptic pyramidal neurons via anterograde and autocrine/paracrine NGF systems.     

The chandelier neuron in schizophrenia

Markers of GABA neurotransmission between chandelier neurons and their synaptic targets, the axon initial segment (AIS) of pyramidal neurons, are altered in the dorsolateral prefrontal cortex (DLPFC) of subjects with schizophrenia. For example, immunoreactivity for the GABA membrane transporter (GAT1) is decreased in presynaptic chandelier neuron axon terminals, whereas immunoreactivity for the GABA(A) receptor α2 subunit is increased in postsynaptic AIS. These alterations are most marked in cortical layers 2-3. In addition, other determinants of the function of chandelier cell-pyramidal neuron synapses, such as ankyrin-G (which regulates the recruitment of sodium channels to the AIS), are also selectively altered in superficial layer pyramidal neurons in subjects with schizophrenia. Each of these components of chandelier cell-pyramidal neuron connectivity exhibits distinctive developmental trajectories in the primate DLPFC, suggesting that disturbances in these trajectories could contribute to the pathogenesis of schizophrenia. Recent findings that inputs from neocortical chandelier neurons are excitatory provide new ideas about the role of this circuitry in the pathophysiology of cortical dysfunction in schizophrenia.     

Microstructure of the neocortex: Comparative aspects

The appearance of the neocortex, its expansion, and its differentiation in mammals, represents one of the principal episodes in the evolution of the vertebrate brain. One of the fundamental questions in neuroscience is what is special about the neocortex of humans and how does it differ from that of other species? It is clear that distinct cortical areas show important differences within both the same and different species, and this has led to some researchers emphasizing the similarities whereas others focus on the differences. In general, despite of the large number of different elements that contribute to neocortical circuits, it is thought that neocortical neurons are organized into multiple, small repeating microcircuits, based around pyramidal cells and their input-output connections. These inputs originate from extrinsic afferent systems, excitatory glutamatergic spiny cells (which include other pyramidal cells and spiny stellate cells), and inhibitory GABAergic interneurons. The problem is that the neuronal elements that make up the basic microcircuit are differentiated into subtypes, some of which are lacking or highly modified in different cortical areas or species. Furthermore, the number of neurons contained in a discrete vertical cylinder of cortical tissue varies across species. Additionally, it has been shown that the neuropil in different cortical areas of the human, rat and mouse has a characteristic layer specific synaptology. These variations most likely reflect functional differences in the specific cortical circuits. The laminar specific similarities between cortical areas and between species, with respect to the percentage, length and density of excitatory and inhibitory synapses, and to the number of synapses per neuron, might be considered as the basic cortical building bricks. In turn, the differences probably indicate the evolutionary adaptation of excitatory and inhibitory circuits to particular functions.     

Spatiotemporal receptive fields: a dynamical model derived from cortical architectonics

We assume that the mammalian neocortex is built up out of some six layers which differ in their morphology and their external connections. Intrinsic connectivity is largely excitatory, leading to a considerable amount of positive feedback. The majority of cortical neurons can be divided into two main classes: the pyramidal cells, which are said to be excitatory, and local cells (most notably the non-spiny stellate cells), which are said to be inhibitory. The form of the dendritic and axonal arborizations of both groups is discussed in detail. This results in a simplified model of the cortex as a stack of six layers with mutual connections determined by the principles of fibre anatomy. This stack can be treated as a multi-input-multi-output system by means of the linear systems theory of homogeneous layers. The detailed equations for the simulation are derived in the Appendix. The results of the simulations show that the temporal and spatial behaviour of an excitation distribution cannot be treated separately. Further, they indicate specific processing in the different layers and some independence from details of wiring. Finally, the simulation results are applied to the theory of visual receptive fields. This yields some insight into the mechanisms possibly underlying hypercomplexity, putative nonlinearities, lateral inhibition, oscillating cell responses, and velocity-dependent tuning curves.     

Intrinsic organization of snake dorsomedial cortex: an electron microscopic and golgi study.

The cellular populations present in dorsomedial cortex in the snakes Constrictor constrictor, Natrix sipendon and Thamnophis sirtalis are described at the light microscopic level using Nissl and Golgi preparations as well as at the ultrastructural level. This area plays a central role in cortical organization in snakes by participating in major commissural and association projections. Systematic analyses of Golgi preparations indicate that five populations of neurons are present in dorsomedial area and have a preferential laminar distribution. Layer 1 stellate cells have somata positioned in the center of the outermost cortical layer, layer 1. Their dendrites are confined to this layer. Double pyramidal cells have their somata loosely packed in layer 2. Their dendrites bear a moderate population of spines, ascending through layer 1 to the pial surface and descending partially through layer 3. Some double pyramidal cells have somata displaced downwards into the upper third of layer 3. These neurons closely resemble the layer 2 double pyramidal cells. Layer 3 stellate cells have somata positioned in the middle third of layer 3. Their dendrites extend in all directions throughout layer 3 and through layer 2 into layer 1. Finally, horizontal cells have their somata positioned deep in layer 3, near the ventricle, and dendrites aligned concentric with the ventricle. Comparison of the organization of the known afferents to dorsomedial area with the distribution of the five cell types suggests that the laminations of both afferent fibres and dorsomedial neurons places specific neuronal populations in synaptic contact with specific sets of afferents.     

Neuronal composition and interneuronal connection of area 5 in the cat parietal association cortex

Area 5 of the cat cortex was studied by Nissl's method and by Golgi's chromate-silver impregnation method. Its typical six-layered structure with well-developed layers of pyramidal cells was revealed. The characteristic features of area 5 are: predominance of pyramidal cells in layers II–III and the presence of large forms (40×26 µ) among them (in layer III); giant pyramidal neurons (70×23 µ) arranged singly or nidally in layer V; large (diameter 25–30 µ) and giant (diameter 40–45 µ) stellate cells with radial dendrites, arranged singly or in groups in layers V–VI; infrequent efferent fusiform neurons (40×20 µ) in layers V–VI. Stellate cells connecting pyramidal neurons in the same or in different layers were found in layers II–VI. Some stellate cells in layers II–III form long horizontal connections within area 5. Interneuronal connections are effected by axosomatic and axodendritic terminals, the latter being more numerous; Dendrodendritic and axoaxonal synapses are less common.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 11, No. 1, pp. 35–42, January–February, 1979.     

Dopamine induces release of calcium from internal stores in layer II lateral entorhinal cortex fan cells.

The entorhinal cortex plays an important role in temporal lobe processes including learning and memory, object recognition, and contextual information processing. The alteration of the strength of synaptic inputs to the lateral entorhinal cortex may therefore contribute substantially to sensory and mnemonic functions. The neuromodulatory transmitter dopamine exerts powerful effects on excitatory glutamatergic synaptic transmission in the entorhinal cortex. Interestingly, inputs from midbrain dopamine neurons appear to specifically target clusters of excitatory cells located in the superficial layers of the entorhinal cortex. We have previously demonstrated that dopamine facilitates synaptic transmission through the activation of D₁-like receptors. This facilitation of synaptic transmission is dependent on both activation of classical D₁-like-receptors, and upon activation of dopamine receptors linked to increases in phospholipase C, inositol triphosphate (IP₃), and intracellular calcium. In the present study we combined electrophysiological recordings of evoked excitatory postsynaptic currents with imaging of intracellular calcium using the fluorescent indicator fluo-4 to monitor calcium transients evoked by dopamine in electrophysiologically identified putative fan and pyramidal cells of the lateral entorhinal cortex. Bath application of dopamine (1 μM), or the phosphatidylinositol (PI)-linked D₁-like-receptor agonist SKF83959 (5 μM), induced reliable and reversible increases in fluo-4 fluorescence and excitatory postsynaptic currents in fan cells, but not in pyramidal cells. In contrast, application of the classical D₁-like-receptor agonist SKF38393 (10 μM) did not result in significant increases in fluorescence. Blocking release of calcium from internal stores by loading cells with the IP₃ receptor blocker heparin (1 mM) or the ryanodine receptor blocker dantrolene (20 μM) abolished both the calcium transients and the facilitation of evoked synaptic currents induced by dopamine. Dopamine also induced calcium transients in fan cells when calcium was excluded from the extracellular medium, further indicating that the calcium transients are linked to release from internal stores. These results indicate that following D₁-like-receptor binding, dopamine selectively induces transient elevations in intracellular calcium via activation of IP₃ and ryanodine receptors, and that these elevations are linked to the facilitation of synaptic responses in putative layer II entorhinal cortex fan cells.