Elementary transcallosal connections of the rabbit sensomotor cortex]

Antidromic responses of two callosal neurones to a local electrical stimulation of the rabbit sensorimotor cortex may be recorded simultaneously with one microelectrode in the homotopic cortical area. In such recording conditions the relative amplitude of extracellularly recorded action potentials of the two neurones is determined primarily by the distance between these neurones and the electrode's tip. In response to the stimulation of the symmetrical area transcallosal monosynaptic excitation of the callosal neurone may occur; two callosal neurones may exite monosynaptically one and the same recorded neurone. The results suggest the existence of clusters or columns, formed jointly by the bodies and terminals of callosal neurones; a functional interconnection between symmetrical clusters or columns may exist, in particular a positive feedback.     

Proton Nuclear Magnetic Resonance Spectroscopy of Primary Cells Derived from Nervous Tissue

Abstract: Cell culture techniques, high-resolution in vitro ¹H nuclear magnetic resonance (NMR) spectroscopy, and chromatographic analyses were used to compare the properties of purified cell populations derived from the PNS and cortical neurones. Cell cultures were immunocytochemically characterised with specific antibodies to ensure purity of the individual cultures. Spectra of perchloric acid extracts of cultured Schwann cells, perineural fibroblasts, dorsal root ganglion neurones, and cortical neurones displayed several common features. However, statistically significant differences were found by ¹H NMR spectroscopy in most metabolites among the cell types studied. In addition, cells could be distinguished by the presence or absence of certain amino acids. For example, N -acetylaspartate was present in dorsal root ganglion neurones and cortical neurones, γ-aminobutyric acid was present in large amounts in cortical neurones, and Schwann cell spectra displayed a large signal from glycine. These results extend our earlier findings that different cell types of the CNS exhibit highly characteristic metabolite profiles to now include the major cell types of the PNS. These latter cell types also exhibit characteristic metabolite compositions, such that even Schwann cells and oligodendrocyte type 2 astrocyte (O-2A) progenitor cells—precursors of the myelinating cells of the CNS and PNS, respectively—can be readily distinguished from each other.     

The organization of double bouquet cells in monkey striate cortex

In previous publications we proposed a model of cortical organization in which the pyramidal cells of the cerebral cortex are organized into modules. The modules are centred around the clusters of apical dendrites that originate from the layer 5 pyramidal cells. In monkey striate cortex such modules have an average diameter of 23 μm and the outputs originating from the modules are contained in the vertical bundles of myelinated axons that traverse the deeper layers of the cortex. The present study is concerned with how the double bouquet cells in layer 2/3 of striate cortex relate to these pyramidal cell modules. The double bouquet cells are visualized with an antibody to calbindin, and it has been shown that their vertically oriented axons, or horse tails, are arranged in a regular array, such that there is one horse tail per pyramidal cell module. Within layer 2/3 the double bouquet cell axons run alongside the apical dendritic clusters, while in layer 4C they are closely associated with the myelinated axon bundles. However, the apical dendrites are not the principal targets of the double bouquet cell axons. Most of the neuronal elements post-synaptic to them are the shafts of small dendrites (60%) and dendritic spines, with which they form symmetric synapses. This regular arrangement of the axons of the double-bouquet cells and their relationship to the components of the pyramidal cells modules supports the concept that there are basic, repeating neuronal circuits in the cortex.     

Precursor cell types in the germinal zone of the cerebral cortex

Retroviral lineage tracing experiments suggest that the cortical ventricular zone is composed of a mixture of precursor cell types. The majority generate a single cell type (neurones, astrocytes or oligodendrocytes) and the remainder generate neurones and a single type of glial cell. Pluripotential precursor cells, that have the ability to generate all three cell types, are not observed. A recent paper, however, reports that when single ventricular zone cells are cultured in isolation, a small percentage of these cells are pluripotential⁽¹⁾. This review will discuss what this knowledge tells us about cortical development.     

Glutamate-mediated overexpression of CD38 in astrocytes cultured with neurones

Recently, a new system of astrocyte-neurone glutamatergic signalling has been identified. It is started in astrocytes by ectocellular, CD38-catalysed conversion of NAD(+) to the calcium mobilizer cyclic ADP-ribose (cADPR). This is then pumped by CD38 itself into the cytosol where the resulting free intracellular Ca(2+) concentration [Ca(2+)](i) transients elicit an increased release of glutamate, which can induce an enhanced Ca(2+) response in neighbouring neurones. Here, we demonstrate that co-culture of either cortical or hippocampal astrocytes with neurones results in a significant overexpression of astrocyte CD38 both on the plasma membrane and intracellularly. The causal role of neurone-released glutamate in inducing overexpression of astrocyte CD38 is demonstrated by two observations: first, in the absence of neurones, induction of CD38 in pure astrocyte cultures can be obtained with glutamate and second, it can be prevented in co-cultures by glutamate receptor antagonists. The neuronal glutamate-mediated effect of neurones on astrocyte CD38 expression is paralleled by increased intracellular cADPR and [Ca(2+)](i) levels, both findings indicating functionality of overexpressed CD38. These results reveal a new neurone-to-astrocyte glutamatergic signalling based on the CD38/cADPR system, which affects the [Ca(2+)](i) in both cell types, adding further complexity to the bi-directional patterns of communication between astrocytes and neurones.     

Neomycin and ruthenium red antagonism of monoaminergic depression of cerebral cortical neurones

Neomycin and ruthenium red, two agents which are known to interfere with the transport and binding of calcium, antagonize the depressant actions of noradrenaline and 5-hydroxytryptamine on cerebral cortical neurones. Ouabain and sodium azide, metabolic inhibitors, also block the action of noradrenaline. These findings support earlier suggestions that monoaminergic depression of cortical neurones involves a calcium-dependent mechanism which may be linked to a membrane ion pump.     

The role of calcium in monoamine-induced depression of cerebral cortical neurones

The calcium antagonists, lanthanum, verapamil and manganese, have been shown to antagonize the depressant actions of ionto-phoretically applied monoamines (noradrenaline, 5-hydroxytryptamine, dopamine and histamine), but not of γ-aminobutyric acid, on cerebral cortical neurones. Cocaine and ethanol, which also affect membrane calcium fluxes, have a similar antagonistic action. These findings suggest that calcium ions are essential for, and perhaps mediate, the characteristic depressant actions of the monoamines on cerebral cortical neurones.     

A presynaptic component of the action of iontophoretically applied flurazepam on feline cortical neurones.

The effect of iontophoretically applied flurazepam on the spike activity of pericruciate cortical neurones of the cat was studied. Flurazepam increased cortical inhibition produced either by local electrical stimulation (which is known to release gamma-aminobutyric acid (GABA) or by iontophoretically applied GABA. Following intravenous treatment with thiosemicarbazide (a GABA-synthesis inhibitor), flurazepam still augmented the action of GABA but was much less effective on electrically evoked cortical inhibition. These findings suggest that part of the action of flurazepam on inhibitory cortical transmission might be at the presynaptic level.     

HMG-CoA reductase inhibition causes neurite loss by interfering with geranylgeranylpyrophosphate synthesis

To determine whether neurite outgrowth depends upon the mevalonate pathway, we blocked mevalonate synthesis in nerve growth factor-treated PC12 cells or primary cortical neurones with atorvastatin, a 3-hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, and substituted different intermediates of the mevalonate pathway. We show that HMG-CoA reductase inhibition causes a profound reduction of neurite length, neurite loss and ultimatively cell death in undifferentiated and pre-differentiated PC12 cells and also in rat primary cortical neurones. Geranylgeranylpyrophosphate, but not farnesylpyrophosphate, squalene or cholesterol, completely compensated for the lack of mevalonate. Our data indicate that, under HMG-CoA reductase inhibition, geranylgeranylpyrophosphate rather than farnesylpyrophosphate or cholesterol is critical for neurite outgrowth and/or maintenance. Loss of neurites is an early manifestation of various neurodegenerative disorders, and dysfunction of isoprenylation might play a role in their pathogenesis.     

Differential effects of unaggregated and aggregated amyloid β protein (1–40) on K+ channel currents in primary cultures of rat cerebellar granule and cortical neurones

The effects of amyloid beta protein on voltage-gated K(+) channel currents were studied using the whole-cell patch-clamp technique. The 1-40 amino acid form of amyloid beta protein was applied to primary cultures of rat cerebellar granule and cortical neurones for 24 h. Both the unaggregated and aggregated forms of the peptide, which have differing biological activities, were used. In cerebellar granule neurones, 24-h pre-incubation with 1 microM unaggregated amyloid beta protein resulted in a 60% increase in the 'A'-type component of K(+) current. Increased delayed rectifier activity was Cd(2+)-sensitive and was presumed to be secondary to an increase in voltage-gated Ca(2+) channel current activity. Unaggregated amyloid beta protein had no effect on any component of the K(+) channel current in cortical neurones. One micromolar of aggregated amyloid beta protein had no effect on K(+) channel current in either cell type but reduced cell survival within 24 h as measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assays. The unaggregated form of amyloid beta protein had no neurotoxic effects when applied to either neurone type for up to 72 h. These data indicate that the unaggregated, non-pathological form of amyloid beta protein causes changes in the ion channel function of neurones, possibly reflecting a physiological role for the peptide.     

Coding with spike shapes and graded potentials in cortical networks

In cortical neurones, analogue dendritic potentials are thought to be encoded into patterns of digital spikes. According to this view, neuronal codes and computations are based on the temporal patterns of spikes: spike times, bursts or spike rates. Recently, we proposed an 'action potential waveform code' for cortical pyramidal neurones in which the spike shape carries information. Broader somatic action potentials are reliably produced in response to higher conductance input, allowing for four times more information transfer than spike times alone. This information is preserved during synaptic integration in a single neurone, as back-propagating action potentials of diverse shapes differentially shunt incoming postsynaptic potentials and so participate in the next round of spike generation. An open question has been whether the information in action potential waveforms can also survive axonal conduction and directly influence synaptic transmission to neighbouring neurones. Several new findings have now brought new light to this subject, showing cortical information processing that transcends the classical models.     

Glutamate dehydrogenase activity in motor cortex neurons during restoration of disrupted visual function]

The recovery of the visual function of rats throughout two weeks after deprivation period (keeping animals in dark chambers for 8 weeks from their birth) leads to a significant normalization of the activity level of glutomatedehydrogenase in the neurones of the III and V layers of the motor cortex. The changes of the enzyme activity in the neurones are accompanied by a diminution of their sizes. The obtained data together with the results of the previous studies (Busnuk, 1976), suggest that the elimination of the visual impulse activity in the early ontogenesis exerts a specific and reversible influence on the morpho-chemical differentiation of neurones both in the visual and in the motor cortical areas. The functional factors determining the direction of changes in the studied parameters of cortical neurones during deprivation and in the rate of their normalization during recovery of the visual function are discussed.     

Interactions between p-tyramine, m-tyramine, or beta-phenylethylamine and dopamine on single neurones in the cortex and caudate nucleus of the rat

p-Tyramine, applied to cortical and caudate neurones with weak iontophoretic currents (0-10 nA), did not usually cause any alteration of base-line firing rate. However, neuronal responses to dopamine (DA) during such weak applications of p-tyramine were greatly enhanced. Cortical neurone responses to noradrenaline (NA) were similarly potentiated, but both cortical and caudate neurone responses to alpha-aminobutyric acid were unaffected by p-tyramine. In addition, weak background applications of DA which did not affect cell firing rate were also without effect on the neuronal responses to the standard application of DA. The responses of cortical neurones to DA were also potentiated by m-tyramine and beta-phenylethylamine applied with weak cationic currents. The results may suggest that trace amines can enhance NA and DA transmission in the central nervous system.     

Probabilistic Identification of Cerebellar Cortical Neurones across Species

Despite our fine-grain anatomical knowledge of the cerebellar cortex, electrophysiological studies of circuit information processing over the last fifty years have been hampered by the difficulty of reliably assigning signals to identified cell types. We approached this problem by assessing the spontaneous activity signatures of identified cerebellar cortical neurones. A range of statistics describing firing frequency and irregularity were then used, individually and in combination, to build Gaussian Process Classifiers (GPC) leading to a probabilistic classification of each neurone type and the computation of equi-probable decision boundaries between cell classes. Firing frequency statistics were useful for separating Purkinje cells from granular layer units, whilst firing irregularity measures proved most useful for distinguishing cells within granular layer cell classes. Considered as single statistics, we achieved classification accuracies of 72.5% and 92.7% for granular layer and molecular layer units respectively. Combining statistics to form twin-variate GPC models substantially improved classification accuracies with the combination of mean spike frequency and log-interval entropy offering classification accuracies of 92.7% and 99.2% for our molecular and granular layer models, respectively. A cross-species comparison was performed, using data drawn from anaesthetised mice and decerebrate cats, where our models offered 80% and 100% classification accuracy. We then used our models to assess non-identified data from awake monkeys and rabbits in order to highlight subsets of neurones with the greatest degree of similarity to identified cell classes. In this way, our GPC-based approach for tentatively identifying neurones from their spontaneous activity signatures, in the absence of an established ground-truth, nonetheless affords the experimenter a statistically robust means of grouping cells with properties matching known cell classes. Our approach therefore may have broad application to a variety of future cerebellar cortical investigations, particularly in awake animals where opportunities for definitive cell identification are limited.     

Evoked potentials arising from neural population elements excited at different times on a warped surface

Certain types of cortical electrical events are non-propagated so that the associated electric fields must have standing wave characteristics. However, cortical electric events typically are generated by neurone populations which cannot be activated simultaneously on impulse driving. Hence the sum of the standing wave fields due to asynchronous activation of adjoining regions of cortical neurones must give the appearance of a traveling wave. Analysis of cortical waveforms is further complicated by curvature in cortical surfaces. A model is presented that shows the effects of curvature and time lag in activation on the form of the potential at points in space around a laminar array of elements simulating a population of cortical neurones. The results are compared with waveforms evoked by single-shock stimulation of the prepyriform cortex in cats.     

Effect of disulfiram on the interconnected activity of cortical neurons in trained cats

The action of disulfiram on interconnected activity of neurones in the visual and motor cortical areas was studied in cats with food-procuring conditioned responses to light. Multiunit activity was recorded from the areas and, by means of amplitude discrimination, separated into impulse flows. Crosscorrelation analysis of the impulse series was used to reveal the character and temporal parameters of interconnected activities of neurones firing in correlation within the limits both of the same cortical area and of the two different ones. A depressing action was shown of the disulfiram on the food-procuring reaction, accompanied by a decrease of the number of pairs of neurones from the visual and motor cortical areas mostly acting in interconnection, interactions with long time delays being mostly affected. The character of action of neighbouring neurones in the visual and motor cortical areas changed in the same direction, expressed in their firing by a "common source" type. The question is discussed of disulfiram influence on interneuronal connections of both types suggesting a decrease of alimentary motivation as well as disturbance of food-procuring conditioned motor coordination.     

Development of longitudinal patterns in the cerebellum of the chicken (Gallus domesticus): a cytoarchitectural study on the genesis of cerebellar modules.

The cytoarchitecture of the cerebellum has been studied in chicken embryos from day 3-20 using serial sections stained with cresylviolet, haematoxylin-eosin and toluidine blue. Three periods have been distinguished in cerebellar development on a basis of cytoarchitectonic characteristics. Of these periods the middle one, which lasts from the 8th to the 15th day, is marked by two subsequent transient longitudinal cytoarchitectonic patterns in the cortical anlage. The first pattern, which exists between days 8 and 11, consists of 4 longitudinal Purkinje cell clusters (of the first order) at either side of the midline. The second pattern, which is most distinct and complete during embryonic days 12-14, is caused by specific localizations of otherwise few, early inwardly migrating granule cells from the external cerebellar matrix (so-called granule raphes), which pass through the layer of Purkinje cell clusters of the first order and thus subdivide these latter into smaller entities: Purkinje cell clusters of the second order. The number of these latter (6 or 7 and 11 or 12 in the anterior and posterior lobes, respectively) correspond to the number of parasagittal modules, which can be discerned on a basis of the organization of fiber connections of the adult cerebellar cortex. Thanks to this similarity various hypotheses can be formulated concerning the significance of the transient cytoarchitectonic patterns in the primitive cortex for the genesis of the modular organization of the cerebellum.     

Evidence for an ascending inhibitory histaminergic pathway to the cerebral cortex.

Previous observations from our laboratory indicate that metiamide is a specific histamine antagonist in rat cerebral cortex. In view of the recent finding that histamine levels and L-histidine decarboxylase (EC 4.1.1.22) activity in cerebral cortex decrease following disruption of the ipsilateral medial forebrain bundle (MFB), the present investigation was undertaken to examine whether iontophoretically applied metiamide antagonizes the inhibition of deep cerebral cortical neurones produced by stimulation of the MFB. In rats anaesthetized with a mixture of methoxyflurane, nitrous oxide and oxygen, stimulation of the ipsilateral MFB or the cortical surface with iontophoretically applied histamine depressed the firing of cortical neurones. Metiamide antagonized the histamine-induced depression and reduced the duration of inhibition produced by MFB stimulation. However, it did not alter the inhibition induced by the cortical surface stimulation. These results indicate that a histaminergic pathway ascending through the MFB may inhibit rat cerebral cortical neurones.     

Self-organization of orientation sensitive cells in the striate cortex

A nerve net model for the visual cortex of higher vertebrates is presented. A simple learning procedure is shown to be sufficient for the organization of some essential functional properties of single units. The rather special assumptions usually made in the literature regarding preorganization of the visual cortex are thereby avoided. The model consists of 338 neurones forming a sheet analogous to the cortex. The neurones are connected randomly to a retina of 19 cells. Nine different stimuli in the form of light bars were applied. The afferent connections were modified according to a mechanism of synaptic training. After twenty presentations of all the stimuli individual cortical neurones became sensitive to only one orientation. Neurones with the same or similar orientation sensitivity tended to appear in clusters, which are analogous to cortical columns. The system was shown to be insensitive to a background of disturbing input excitations during learning. After learning it was able to repair small defects introduced into the wiring and was relatively insensitive to stimuli not used during training.