Stellate Neurons In Layer Research Articles

Layer 4 pyramidal neuron dendritic bursting underlies a post-stimulus visual cortical alpha rhythm

Alpha rhythms (9–11 Hz) are a dominant feature of EEG recordings, particularly over occipital cortex on cessation of a visual stimulation. Little is known about underlying neocortical mechanisms so here we constructed alpha rhythm models that follow cessation of cortical stimulation. The rhythm manifests following a period of gamma frequency activity in local V1 networks in layer 4. It associates with network level bias of excitatory synaptic activity in favour of NMDA- rather than AMPA-mediated signalling and reorganisation of synaptic inhibition in favour of fast GABAA receptor-mediated events. At the cellular level the alpha rhythm depended upon the generation of layer 4 pyramidal neuron dendritic bursting mediated primarily by PPDA-sensitive NR2C/D-containing NMDA receptors, which lack the magnesium-dependent open channel block. Subthreshold potassium conductances are also critical. The rhythm dynamically filters outputs from sensory relay neurons (stellate neurons in layer 4) such that they become temporally uncoupled from downstream population activity.

Simulation of signal flow in 3D reconstructions of an anatomically realistic neural network in rat vibrissal cortex

The three-dimensional (3D) structure of neural circuits represents an essential constraint for information flow in the brain. Methods to directly monitor streams of excitation, at subcellular and millisecond resolution, are at present lacking. Here, we describe a pipeline of tools that allow investigating information flow by simulating electrical signals that propagate through anatomically realistic models of average neural networks. The pipeline comprises three blocks. First, we review tools that allow fast and automated acquisition of 3D anatomical data, such as neuron soma distributions or reconstructions of dendrites and axons from in vivo labeled cells. Second, we introduce NeuroNet, a tool for assembling the 3D structure and wiring of average neural networks. Finally, we introduce a simulation framework, NeuroDUNE, to investigate structure-function relationships within networks of full-compartmental neuron models at subcellular, cellular and network levels. We illustrate the pipeline by simulations of a reconstructed excitatory network formed between the thalamus and spiny stellate neurons in layer 4 (L4ss) of a cortical barrel column in rat vibrissal cortex. Exciting the ensemble of L4ss neurons with realistic input from an ensemble of thalamic neurons revealed that the location-specific thalamocortical connectivity may result in location-specific spiking of cortical cells. Specifically, a radial decay in spiking probability toward the column borders could be a general feature of signal flow in a barrel column. Our simulations provide insights of how anatomical parameters, such as the subcellular organization of synapses, may constrain spiking responses at the cellular and network levels.

Dorsal–ventral organization of theta‐like activity intrinsic to entorhinal stellate neurons is mediated by differences in stochastic current fluctuations

The membrane potential dynamics of stellate neurons in layer II of the medial entorhinal cortex are important for neural encoding of location. Previous studies suggest that these neurons generate intrinsic theta-frequency membrane potential oscillations, with a period that depends on neuronal location on the dorsal–ventral axis of themedial entorhinal cortex, and which in behaving animals could support generation of grid-like spatial firing fields. To address the nature and organization of this theta-like activity, we adopt the Lombmethod of least-squares spectral analysis. We demonstrate that peaks in frequency spectra that differ significantly from Gaussian noise do not necessarily imply the existence of a periodic oscillator, but can instead arise from filtered stochastic noise or a stochastic random walk. We show that theta-like membrane potential activity recorded fromstellate neurons in mature brain slices is consistentwith stochastic mechanisms, but not with generation by a periodic oscillator. The dorsal–ventral organization of intrinsic theta-likemembrane potential activity, and themodification of this activity during block of HCN channels, both reflect altered frequency distributions of stochastic spectral peaks, rather than tuning of a periodic oscillator. Our results demonstrate the importance of distinguishing periodic oscillations from stochastic processes.We suggest that dorsal–ventral tuning of theta-like membrane potential activity is due to differences in stochastic current fluctuations resulting from organization of ion channels that also control synaptic integration.

Adenosine modulates the excitability of layer II stellate neurons in entorhinal cortex through A1 receptors

Stellate neurons in layer II entorhinal cortex (EC) provide the main output from the EC to the hippocampus. It is believed that adenosine plays a crucial role in neuronal excitability and synaptic transmission in the CNS, however, the function of adenosine in the EC is still elusive. Here, the data reported showed that adenosine hyperpolarized stellate neurons in a concentration-dependent manner, accompanied by a decrease in firing frequency. This effect corresponded to the inhibition of the hyperpolarization-activated, cation nonselective (HCN) channels. Surprisingly, the adenosine-induced inhibition was blocked by 3 μM 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), a selective A(1) receptor antagonists, but not by 10 μM 3,7-dimethyl-1-propargylxanthine (DMPX), a selective A(2) receptor antagonists, indicating that activation of adenosine A(1) receptors were responsible for the direct inhibition. In addition, adenosine reduced the frequency but not the amplitude of miniature EPSCs and IPSCs, suggesting that the global depression of glutamatergic and GABAergic transmission is mediated by a decrease in glutamate and GABA release, respectively. Again the presynaptic site of action was mediated by adenosine A(1) receptors. Furthermore, inhibition of spontaneous glutamate and GABA release by adenosine A(1) receptor activation was mediated by voltage-dependent Ca(2+) channels and extracellular Ca(2+) . Therefore, these findings revealed direct and indirect mechanisms by which activation of adenosine A(1) receptors on the cell bodies of stellate neurons and on the presynaptic terminals could regulate the excitability of these neurons.

Tuning of Synaptic Integration in the Medial Entorhinal Cortex to the Organization of Grid Cell Firing Fields

Neurons important for cognitive function are often classified by their morphology and integrative properties. However, it is unclear if within a single class of neuron these properties tune synaptic responses to the salient features of the information that each neuron represents. We demonstrate that for stellate neurons in layer II of the medial entorhinal cortex, the waveform of postsynaptic potentials, the time window for detection of coincident inputs, and responsiveness to gamma frequency inputs follow a dorsal-ventral gradient similar to the topographical organization of grid-like spatial firing fields of neurons in this area. We provide evidence that these differences are due to a membrane conductance gradient mediated by HCN and leak potassium channels. These findings suggest key roles for synaptic integration in computations carried out within the medial entorhinal cortex and imply that tuning of neural information processing by membrane ion channels is important for normal cognitive function.

The morphological changes of pyramidal and spiny stellate cells in the primary visual cortex of chronic morphine treated cats

Morphine exposure may have a negative effect on the receptive field properties of neurons in primary visual cortex of cats. The present experiment used morphological methods in order to investigate whether chronic morphine treatment also affects dendritic characters of these neurons. According to the Sholl analysis and dendritic branch order analysis, we obtained the dendritic length and calculated the spine density on dendrites of the pyramidal neurons in layer III and the spiny stellate neurons in layer IV. The results showed that morphine exposure induced significant decreases in the total dendritic length and spine density on both pyramidal and spiny stellate neurons. The further branch order analysis revealed that spine density was decreased at every (first to fourth) branch order of dendrites of pyramidal and spiny stellate neurons. Decrease in dendritic length of the pyramidal neurons was observed only at the fourth branch order, while the spiny stellate neurons had shorter dendrite at the second and third branch order. These findings may underlie the degradation of receptive field properties of the primary visual cortex neurons following chronic morphine exposure.

HCN1 Channels Control Resting and Active Integrative Properties of Stellate Cells from Layer II of the Entorhinal Cortex

Whereas recent studies have elucidated principles for representation of information within the entorhinal cortex, less is known about the molecular basis for information processing by entorhinal neurons. The HCN1 gene encodes ion channels that mediate hyperpolarization-activated currents (I(h)) that control synaptic integration and influence several forms of learning and memory. We asked whether hyperpolarization-activated, cation nonselective 1 (HCN1) channels control processing of information by stellate cells found within layer II of the entorhinal cortex. Axonal projections from these neurons form a major component of the synaptic input to the dentate gyrus of the hippocampus. To determine whether HCN1 channels control either the resting or the active properties of stellate neurons, we performed whole-cell recordings in horizontal brain slices prepared from adult wild-type and HCN1 knock-out mice. We found that HCN1 channels are required for rapid and full activation of hyperpolarization-activated currents in stellate neurons. HCN1 channels dominate the membrane conductance at rest, are not required for theta frequency (4-12 Hz) membrane potential fluctuations, but suppress low-frequency (<4 Hz) components of spontaneous and evoked membrane potential activity. During sustained activation of stellate cells sufficient for firing of repeated action potentials, HCN1 channels control the pattern of spike output by promoting recovery of the spike afterhyperpolarization. These data suggest that HCN1 channels expressed by stellate neurons in layer II of the entorhinal cortex are key molecular components in the processing of inputs to the hippocampal dentate gyrus, with distinct integrative roles during resting and active states.

Long-Term Depression in Identified Stellate Neurons of Juvenile Rat Entorhinal Cortex

The entorhinal cortex (EC) serves as a gateway to the hippocampus and plays a pivotal role in memory processing in the brain. Superficial layers of the EC convey the cortical input projections to the hippocampus, whereas deep layers of the EC relay hippocampal output projections back to the superficial layers of the EC or to other cortical regions. Whereas the EC expresses long-term potentiation (LTP) and depression (LTD), the underlying cellular and molecular mechanisms have not been determined. Because the axons of the stellate neurons in layer II of the EC form the perforant path that innervates the dentate gyrus granule cells of the hippocampus, we studied the mechanisms underlying the long-term plasticity in identified stellate neurons. Application of high-frequency stimulation (100 Hz for 1 s, repeated 3 times at an interval of 10 s) or forskolin (50 microM) failed to induce significant changes in synaptic strength, whereas application of pairing (presynaptic stimulation at 0.33 Hz paired with postsynaptic depolarization from -60 to -10 mV for 5 min) or low-frequency stimulation (LFS, 1 Hz for 15 min) paradigm-induced LTD. Pairing- or LFS-induced LTDs were N-methyl-D-aspartate receptor-dependent and occluded each other suggesting that they have the similar cellular mechanism. Pairing-induced LTD required the activity of calcineurin and involved AMPA receptor endocytosis that required the function of ubiquitin-proteasome system. Our study provides a cellular mechanism that might in part explain the role of the EC in memory.

Detection of entorhinal layer II using Tesla magnetic resonance imaging

The entorhinal cortex lies in the mediotemporal lobe and has major functional, structural, and clinical significance. The entorhinal cortex has a unique cytoarchitecture with large stellate neurons in layer II that form clusters. The entorhinal cortex receives vast sensory association input, and its major output arises from the layer II and III neurons that form the perforant pathway. Clinically, the neurons in layer II are affected with neurofibrillary tangles, one of the two pathological hallmarks of Alzheimer's disease. We describe detection of the entorhinal layer II islands using magnetic resonance imaging. We scanned human autopsied temporal lobe blocks in a 7T human scanner using a solenoid coil. In 70 and 100 microm isotropic data, the entorhinal islands were clearly visible throughout the anterior-posterior extent of entorhinal cortex. Layer II islands were prominent in both the magnetic resonance imaging and corresponding histological sections, showing similar size and shape in two types of data. Area borders and island location based on cytoarchitectural features in the mediotemporal lobe were robustly detected using the magnetic resonance images. Our ex vivo results could break ground for high-resolution in vivo scanning that could ultimately benefit early diagnosis and treatment of neurodegenerative disease.

Coincidence detection and changes of synaptic efficacy in spiny stellate neurons in rat barrel cortex.

Paired whole-cell voltage recordings were made from synaptically connected spiny stellate neurons in layer 4 of the barrel field in young (P14) rat somatosensory cortex. When postsynaptic action potentials (APs) followed each of 5 presynaptic APs in a 10- or 20-Hz train by less than 25 ms, subsequent unitary EPSP amplitudes were persistently reduced. Induction of long-term depression (LTD) depended on activation of group II metabotropic glutamate receptors, but not on NMDA or AMPA receptors. Reducing postsynaptic increases in intracellular calcium ([Ca2+]i) by intracellular loading with a fast- (BAPTA) or a slow- (EGTA) acting Ca2+ buffer blocked synaptic depression. Analysis of EPSP failures suggested mediation of LTD by a reduction in release probability. We propose a mechanism by which coincident activity results in long-lasting reduction of synaptic efficacy between synaptically connected neurons.

Polyneuronal innervation of spiny stellate neurons in cat visual cortex.

Our hypothesis was that spiny stellate neurons in layer 4 of cat visual cortex receive polyneuronal innervation. We characterised the synapses of four likely sources of innervation by three simple criteria: the type of synapse, the target (spine, dendritic shaft), and the area of the presynaptic bouton. The layer 6 pyramids had the smallest boutons and formed asymmetric synapses mainly with the dendritic shaft. The thalamic afferents had the largest boutons and formed asymmetric synapses mainly with spines. The spiny stellates had medium-sized boutons and formed asymmetric synapses mainly with spines. We used these to make a "template" to match against the boutons forming synapses with the spiny stellate dendrite. Of the asymmetric synapses, 45% could have come from layer 6 pyramidal neurons, 28% from spiny stellate neurons, and 6% from thalamic afferents. The remaining 21% of asymmetric synapses could not be accounted for without assuming some additional selectivity of the presynaptic axons. Additional asymmetric synapses may come from a variety of sources, including other cortical neurons and subcortical nuclei such as the claustrum. Of the symmetric synapses, 84% could have been provided by clutch cells, which form large boutons. The remainder, formed by small boutons, probably come from other smooth neurons in layer 4, e.g., neurogliaform and bitufted neurons. Our analysis supports the hypothesis that the spiny stellate receives polyneuronal innervation, perhaps from all the sources of boutons in layer 4. Although layer 4 is the major recipient of thalamic afferents, our results show that they form only a few percent of the synapses of layer 4 spiny stellate neurons.

Distribution and morphology of area 17 neurons that project to the cat's extrastriate cortex

We used intracellular dye injections in lightly fixed cortical slices to examine the distribution and morphology of area 17 neurons that project to extrastriate cortex. Both the projection to the medial bank of the posterior lateral suprasylvian sulcus (PMLS) and the projection to area 18 arise from a number of different morphological types distributed throughout layers 2-6. The majority are found in the superficial layers and include large, medium, and small pyramidal neurons. Some are also found in the deep layers and they include very large, pyramidal neurons as well as some heretofore undescribed, fusiform neurons. The projection to area 18 contains two types not found in the projection to PMLS: spinous stellate neurons in layer 4 and inverted pyramidal neurons in layer 3. Finding that a variety of morphological types contribute projections to a single cortical area raises the possibility that corticocortical projections, like retinogeniculate and geniculocortical projections, comprise multiple parallel pathways with different physiological properties and patterns of termination. Finding that the projection to area 18 contains morphological types that do not project to PMLS indicates that the projections from area 17 are likely to contribute to the functional specialization of extrastriate visual areas.

Neuronal Development in Vitamin B6 Deficiency

The morphological changes observed in developing brain regions associated with maternal vitamin B6 deficits are summarized in Table 4. Brain development is a complex and orderly process consisting of cell division, proliferation, migration, and maturation. In the rat, vitamin B6 deficits imposed in utero and up to 30 days postnatal interfere with this orderly process. Deficits of the vitamin imposed in utero have been associated with reduced numbers of total and normal neurons in neocortex and with increased shrunken neurons (700-1500% of controls) in this region. These changes reflect the critical role of vitamin B6 in both neurogenesis and neuron longevity in neocortex. Postnatal cellular events in the neocortex, that is, neuron differentiation and synaptogenesis, were also altered by vitamin B6 deficits; higher order dendrites were reduced on stellate neurons in Layer II and on pyramidal neurons in Layer V. Synaptic density was less in the neutrophil of neocortex and in caudate/putamen, but structural integrity of the synapse was maintained. In cerebellum, both the molecular and granular areas were reduced, the monolayer organization of Purkinje cells was disrupted, and dendritic arborization of the cells was decreased. The number of myelinated axons, as determined by electron microscopy, was decreased in the mediodorsal portion of the pyramidal tract in the medulla oblongata as well as the specific activity of myelination of the total brain. Thus the functional consequences of vitamin B6 deficits during neuronal development may be through reduced connections among neurons and decreased myelination, which alter the rate and magnitude of transmission of nerve impulses.

Effects of Maternal Restriction in Vitamin B-6 on Neocortex Development in Rats: Neuron Differentiation and Synaptogenesis

Effects of maternal restrictions in vitamin B-6 on neuron differentiation and synaptogenesis in developing neocortex were examined. Rats were fed ad libitum a vitamin B-6-free diet supplemented with 0.0 or 0.6 mg pyridoxine hydrochloride (PN.HCl)/kg diet during gestation followed by a control level of 7.0 mg/kg diet during lactation, or they were fed the vitamin B-6-free diet supplemented with 0.6 or 7.0 mg PN.HCl/kg diet throughout gestation and lactation. Neocortices of the offspring were examined at 30 d of age by light and electron microscopy. All maternal restrictions in vitamin B-6 reduced the number of higher order dendrites on stellate neurons in layer II and on pyramidal neurons in layer V of the neocortex and decreased synaptic density in the neuropil of the neocortex. The findings indicated that vitamin B-6 restriction during gestation, either marginal or severe, was the critical treatment factor that adversely affected synaptogenesis and at least one event in neuron differentiation in the neocortex, the arborization of dendrites.

Structural alterations of dendritic spines induced by neural degeneration of their presynaptic afferents.

Morphological parameters were compared for dendritic spines of spiny stellate neurons in layer IV of the barrel region of mouse somatosensory cortex, which synapse with degenerated thalamocortical afferents (TC spines) and with intact, unidentified axon terminals (UI spines). Spiny stellate neurons were labeled for light and electron microscopic identification by Golgi impregnation and gold toning. Dendritic spines were examined in series of thin sections, and TC spines were ultrastructurally detectable because of the degeneration-induced characteristic appearance of the TC axon terminals. Results show that the means of the width of the spine head and of the length of the spine stalk were significantly higher in TC spines than in UI spines by about 11 and 25%, respectively. The variability of these two morphological parameters was significantly lower for TC spines. The mean of the spine stalk width at the narrowest cross section of the stalk was about 0.12 microns, with no significant difference observed between the two spine groups. No specific relationship was found in either the TC or the UI groups of spines between the length of the spine stalk and the width of the spine stalk at its narrowest profile. As structural features typifying transneuronal degeneration were not observed along the dendritic spines examined, it is speculated that the morphological differences encountered between the TC and UI spines may result, at least in part, from the degeneration-induced synaptic inactivity of the TC axospinous synapses, rather than exclusively from any direct effects of the degeneration process.

The spiny stellate neurons in layer IV of the human auditory cortex. A Golgi study

The spiny stellate neurons have been studied by the Golgi method in the auditory koniocortex and parakoniocortex of man. Spiny stellate cells are a consistent though not very common component of layer IV. They are not confined to specific sublayers but occur at all depths of layer IV, and also in layer IIIc. Spiny stellate cells in the auditory areas show a great variety of their dendritic arborization pattern. The presence of all intermediate forms between small pyramidal cells—which constitute the dominant cell type in layer IV and which display an extraordinary heteromorphism—and spiny stellate cells shows the close kinship between both neuronal types. The morphology and distribution of spines along the dendrites of spiny stellate neurons are similar to those of the small pyramidal cells of the same layer. The axons, which were impregnated only in their proximal portions, mostly descend, giving rise to recurrent ascending collaterals, but initially ascending axons do also occur. Spiny stellate neurons are present in the different cytoarchitectonic areas examined, and thus they are not confined to the auditory koniocortex.

Quantification of thalamocortical synapses with spiny stellate neurons in layer IV of mouse somatosensory cortex.

The distribution of thalamocortical (TC) and other synapses involving spiny stellate neurons in layer IV of the barrel region of mouse primary somatosensory cortex (SmI) was examined in seven male CD/1 mice. TC axon terminals were labeled by lesion-induced degeneration, which has been shown to label reliably all TC synapses in mouse barrel cortex. Spiny stellate neurons, labeled by Golgi impregnation and gold toning, were identified with the light microscope prior to thin sectioning and electron microscopy. Analysis of eight dendritic segments from seven spiny stellate neurons showed that most of their synapses are with their dendritic spines, rather than with their shafts. Axospinous synapses are primarily of the asymmetrical type, whereas axodendritic synapses are mainly of the symmetrical type. Dendrites of spiny stellate neurons consistently form thalamocortical synapses, most of which involve spine heads rather than spine stalks or dendritic shafts. From 10.4% to 22.9% of all asymmetrical synapses with dendrites of spiny stellate neurons involve TC axon terminals. In general, this is a higher range than the ranges that characterize the TC synaptic connectivity of dendrites belonging to other types of neurons, implying that spiny stellate neurons are perhaps more strongly influenced by TC synaptic input than other types of cortical neurons examined previously. Spines involved in TC synapses were distributed irregularly along each of the stellate cell dendrites; about half of the interspinous intervals between these spines were about 5 microns or less. Modulations of the efficacy of TC synaptic input to dendrites of layer IV spiny stellate neurons are discussed in the light of recently reported computer simulated analyses of axospinous synaptic connections.

Distribution and ultrastructure of thalamic afferents in the cat auditory cortex

Ultrastructural features of thalamic afferent fibers were studied in the cat auditory cortex in the early stages (on the 4th day) of experimental degeneration produced by destruction of the medial geniculate body. A coordinate grid was used in conjunction with an electron micro-scope to study the topography of the degenerating elements over wide areas of sections, so that the density of degeneration could be determined quantitatively in different layers of the cortex. Degenerating axons were found in all layers. Most of the large (5–7 µ) degenerating axons are located in layer VI; their diameters were smaller in the upper layers of the cortex. Degenerative changes affecting synaptic terminals of thalamo-cortical afferents were of the "dark" type. Fibers of the geniculo-cortical tract were shown to terminate mainly in cortical layer IV. A few degenerating synapses were found in the molecular layer. Terminals with sperical synaptic vesicles are found mainly on the spines of dendrites where they form "asymmetrical" contacts. A few degenerating axo-somatic synapses were observed on stellate neurons in layer IV. The results are discussed in connection with electrophysiological investigations of the cat auditory cortex during stimulation of specific afferent fibers.