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[Reflection of the plastic properties of 2 neurons with a common monosynaptic input in the statistical characteristics of their impulse activity].

Changes of cross-correlation histograms (CCH) of impulse trains and of mean interspike intervals (ISI) of neurones N1 and N2 with a common monosynaptic excitatory or inhibitory-excitatory input from N3, at changes of efficiency of interneuronal connections, neurone excitability and summate action on them of independent random afferent synaptic inflows were studied by methods of mathematical and biomathematical modelling of neuronal interaction. It was shown that the increase of amplitude of the central peak (trough) of a normalized CCH of N1-N2 accompanied by reduction of mean ISI of N1 and N2, is either a sign of an increase of the amplitude of postsynaptic potentials of N1 and N2 elicited by impulses of the nonrecorded N3 or a sign of an increase of mean ISI of N3.

Action Potentials↗

Mapping of the distribution of polysialylated neural cell adhesion molecule throughout the central nervous system of the adult rat: an immunohistochemical study.

In the nervous system, the neural cell adhesion molecule changes at the cell surface during development, from a form highly enriched in polysialic residues to several isoforms containing much less sialic acid, and is thought to participate in the structuring of neuronal groups and in the establishment of neuronal connections. Recent observations have indicated, however, that it may not be restricted to developing tissues since it is still present in certain adult neuronal centres which can undergo morphological reorganization. In this study, therefore, we examined systematically the distribution of polysialylated neural cell adhesion molecule immunoreactivity throughout the central nervous system of adult male and female rats, using light microscopic immunocytochemistry and immunoblot analysis with an antibody that specifically recognizes the polysialic residues of the molecule. Concomitantly, we compared this immunoreactivity to that due to all isoforms of the neural cell adhesion molecule, detected with a polyclonal serum raised against the NH2-terminal of the protein. Immunoreactivity due to the polysialylated isoform was consistently visualized in several discrete areas of the adult brain and spinal cord. An intercellular punctate immunolabelling characterized the staining in certain hypothalamic and thalamic nuclei, superficial laminae of the dorsal horn of the spinal cord, ventral portion of the dentate gyrus of the hippocampus, lateral geniculate, parabrachial and habenular nuclei, bed nucleus of the stria terminalis, mesencephalic central gray and olfactory bulb. In other areas, such as the piriform cortex, dorsal aspect of the dentate gyrus and fimbria and lamina X of the spinal cord, isolated neuronal-like cells were either completely filled with immunolabel or showed a surface reaction on their cell bodies and processes. Highly immunoreactive isolated glial-like cells were also noted within the ependymal layer of the central canal and lateral ventricles and at times in the peripheral white matter of the spinal cord. In contrast to this discrete localization, staining due to all isoforms of the neural cell adhesion molecule was widespread and diffuse throughout the brain and spinal cord. The expression of the polysialylated isoform in the supraoptic nucleus and hippocampus was confirmed by immunoblot analysis; it occurred together with weakly sialylated isoforms. No obvious differences were detected in the amount or distribution of immunoreactivity due to the polysialylated isoform in relation to the sex or age of the animals (between three and 12 months of age). Our study thus demonstrates that well-defined areas of the central nervous system of the adult rat continue to express the polysialylated isoform of the neural cell adhesion molecule.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Ontogenetic studies of seizure patterns and seizure activities induced by cortical focus.

Ontogenetic studies of epileptogenic process were carried out in albino rats ranging in age from birth to 45 days. Experimental epilepsy was produced by two different procedures and the results were compared with each other. Tungstic acid gel was applied to the motor area of the left side of the cortex, and the following results were obtained. The latency of the seizure appearance was long during 10 days after birth, became progressively short thereafter and reached the minimum in about 20 days of age, and gradually returned to the adult level again by 45 days of age. No abvvious seizure was exhibited until five days of age. Seizure patterns developed from tonic or twitch-like jerky convulsion (10 days old) to rhythmic or clonic type of seizure (13 days old), and the seizure patterns similar to those in the adult rat were observed in about 20 days of age. Cortical seizure activity was initially observed in about 10-day-old rats; single high amplitude slow wave appeared and small spikes became superimposed on it in the course of maturation. Atypical spike and wave complexes were observed after 20 days of age. Electrical stimulation was applied to the left cortical motor area by constant current stimulator, and the following seizure patterns were observed: No obvious seizure could be elicited in newborn rat, whereas from three days of age, tonic seizure of the whole body, and from seven days old twitch-like convulsion of extremities were observed. In ages from 10 to 20 days, seizure induced by electrical stimulation was mainly tonic in pattern; extension of forelimbs and flexion of hindlimbs in most cases were observed before 13 days old, but both fore-and hindlimbs were extended therafter. Tonic-clonic seizure patterns were exhibited after 20 days of age. From these results, it was considered that tonic convulsions and high voltage slow cortical seizure activites were produced from the activites of the local cortical neuronal connections, and rhythmic and/or clonic seizure patterns and spike and wave seizure activities were elicited from the more complex, i.e. cortico-subcortical neuronal circuits. Possible contributing factors for the determination of seizure susceptibility in immautre rats were also discussed.

Age Factors↗

Dual intracellular recording of neocortical neurons in a neuron-glia co-culture system.

Measurement of synchronized activities in neocortical networks is critical for understanding integrated brain function. We describe here a procedure for intracellular recording of activity in two functionally connected neurons using patch electrodes in neuron-glia co-cultures. Rat neocortical neurons survive on monolayers of astrocytes for far longer periods (3-4 weeks) than in their absence. This prolonged survival facilitates the detection of functional synapses which are readily identified in vitro by means of dual intracellular recording using two patch electrodes. The neuron-glia co-culture system appears to be useful not only for studying the formation of neural networks in vitro but also for determining the effects of physiologically active substances and chemicals on synapse formation.

Animals↗

Specific and nonspecific multiple unit activities during the onset of pentylenetetrazol seizures. II. Acute lesions interrupting nonspecific system connections.

Nonspecific cortical, thalamic, mesencephalic, and pontine multiple unit activities (MUA) and changes in EEG and MUS of the sciatic nerve after threshold pentylenetetrazol activation were studied in three groups of animals in which neuronal connections were interrupted at three different levels of the central nervous system: spinal, mesencephalic, and prethalamic. Maximal increments of nonspecific MUA and maximal increments and maximal decrements of sciatic MUA after pentylenetetrazol from each group of lesioned animals were statistically compared with tose observed in intact animals. 1. Pentylenetetrazol threshold for producing cortical tonic-clonic EEG discharges was increased in animals with nesencephaic and prethalamic lesions but was not modified in animals with spinal transection. 2. Cortical MUA maximal increment was significantly decreased in mesencephalic and prethalamic lesioned animals, whereas thalamic MUA maximal increment was significantly decreased in mesencephalic and significantly increased in prethalamic lesioned animals. Pontine MUA maximal increment was significantly increased in spinal, mesencephalic, and prethalamic lesioned animals, and mesencephalic M-A maximal increment was not significantly modified in either prethalamic lesioned or in spinal transected animals. 3. Sciatic MUA maximal increment and maximal decrement were significatly decreased in spinal transected animals, whereas only maximal increment was significantly decreased in mesencephalic and only maximal decrement was significantly decreased in prethalamic lesioned animals. These results based on lesion experiments permit us to infer than under normal cinditions the development of generalized seizures induced by threshold pentylenetetrazol injection is highly dependent upon the neuronal interactions between nonspecific structures at different levels of the central nervous system. The possible nature of these neuronal interactions in the intact animals is discussed.

Animals↗

[Neurobiology of learning--the basis of an alteration process].

In contrast to the opinion that prevailed in the 1980es, there is now increasing evidence that the plasticity of the human brain, i. e. its remarkable ability to adapt to and change with experience, is, under normal conditions, a lifelong phenomenon. Representations of the environment are associated with activations and biochemical modifications in neuronal networks, which will be stabilized, modified or will wither in the course of cumulated experience. The capability to modify the biochemistry of synapses as well as the growth and change in terms of rewiring of synapses, dendritic branching and glial cell proliferation via the dialogue of synapses and genes, results in specific changes in neuronal connectivity and function. On the neurotransmitter level, glutamate and gamma-amino-butyric acid (GABA) as well as dopamine and serotonin, but also endorphin and encephalin, have a key position in this context. These neurotransmitter systems modulate neuronal plasticity on the neuronal level; on the behavioural level they influence affect, emotion, positive motivation and the correct evaluation of environmental stimuli. Experience, action as well as learning and memory are influenced by these systems. A basic thesis of this paper is that these mechanisms are involved in neuronal plasticity and that learning and memory are thus not only used and reused in structuring the CNS during the initial establishment of connections in the immature brain, in lifelong memory consolidation or the rewiring after brain damage, but can also be used to mould experience, learning und behaviour during psychotherapy and rehabilitation in adults.

Brain↗

Mapping and ultrastructure of serotonin-immunoreactive neurons in the optic lobes of three insect species.

With antibodies to serotonin (5-HT) we have mapped immunoreactive neurons in the optic lobes of three species, the blowfly Calliphora, the desert ant Cataglyphis, and the worker bee Apis. The main emphasis in this investigation is on a system of 5-HT-positive neurons connecting the most peripheral neuropil of the optic lobes, the lamina, to more central neuropil regions. To aid in electron microscopical identification of these neurons we used immunocytochemistry at the EM-level and Golgi-EM for Calliphora and horseradish peroxidase (HRP) labelling for the other two insects. The immunoreactive terminals in Calliphora and the HRP-labelled ones in the other insects contain large (c. 100 nm) granular vesicles and smaller (c.60 nm) clear vesicles. In Cataglyphis and Apis the profiles with granular vesicles are presynaptic to second order neurons of the lamina, whereas in Calliphora no synaptic contacts were found. In this animal the 5-HT-positive terminals are situated distal to the synaptic layer of the lamina, in a region of retinal photoreceptor axons and perikarya of the lamina monopolar neurons. In Catagylphis and Apis the interactions of the 5-HT-neurons with the laminar neurons might occur through chemical synapses, whereas in Calliphora neuroactive substance could be released non-synaptically from varicosities distal to the synaptic layer. The possible involvement of 5-HT in control of neuronal activity in the optic lobes is discussed.

Animals↗

Pyramidal neurons of the rat cerebral cortex, immunoreactive to nicotinic acetylcholine receptors, project mainly to subcortical targets.

Cortical neurons immunoreactive to nicotinic acetylcholine receptors (nAChR) of the rat brain were characterized with monoclonal antibodies directed to ACh-binding subunits (alpha 4) or to ACh-structural subunits (beta 2). A heterogeneous population of nAChR-LI neurons was found in all cortical regions. The most prominent immunoreactive neurons were pyramids of layers V and II-III. The nonpyramidal positive neurons were fusiform horizontally oriented neurons of layer VIb, small cells of layer I and round or ovoid neurons of layers II-V. Double labeled experiments (immunohistochemistry and fluorescent retrograde tracers) showed that cholinoceptive pyramidal neurons of layer V project mainly to subcortical targets such as caudate-putamen, superior colliculus, and pontine nuclei, while very few nAChR positive neurons connect to other cortical areas. These findings suggest that the mainly excitatory effect that has been attributed to the cholinergic innervation upon the cortical neurons may have a greater influence upon the cortico-subcortical output than the corticortical one.

Animals↗

Cytoskeletal dynamics underlying collateral membrane protrusions induced by neurotrophins in cultured Xenopus embryonic neurons.

The establishment and refinement of neuronal connections depend on dynamic modification of the morphology and physiology of developing axons in response to extrinsic factors. In embryonic cultures of Xenopus spinal neurons, acute application of brain-derived neurotrophic factor (BDNF) induced rapid collateral protrusion of filopodium-like microspikes and lamellipodia along the neurite processes, leading to a morphologic alternation of the neuron. Both types of membrane protrusions contained high concentrations of actin filaments and depended on the polymerization of the actin cytoskeleton. Immunofluorescent staining, however, revealed the presence of microtubules (MTs) in lamellipodia induced by BDNF. These MTs appeared to have arisen from debundling of MTs in the neurite shaft at the protrusion sites, splaying and extending in the rapidly protruding lamellipodia. Inhibition of microtubule polymerization by nocodazole largely abolished the formation of lamellipodia but not of microspikes. Taken together, our results suggest that collateral sprouting of microspikes and lamellipodia involve distinctly different cytoskeletal mechanisms. Although the actin cytoskeleton is solely responsible for microspike formation, cooperative efforts by microtubules and actin filaments are essential for lamellipodial protrusion in response to extrinsic factors.

Actin Cytoskeleton↗

Signaling at the vertebrate synapse: new roles for embryonic morphogens?

The formation of synapses is critical for functional neuronal connectivity. The coordinated assembly at both sides of the synapse is fundamental for the proper apposition of the neurotransmitter release machinery on the presynaptic neuron and the clustering of neurotransmitter receptors and ion channels on the receptive postsynaptic cell. This process requires bidirectional communication between the presynaptic neuron and its postsynaptic target, another neuron, or muscle fiber. Extracellular signals such as WNT, TGF-beta, and FGF factors are emerging as key target-derived signals required for the initial stages of synaptic assembly. Studies in invertebrates are also providing new insights into the function of these signals in synaptic growth and homeostasis. During early embryonic patterning, WNT, TGF-beta, and FGF factors function as typical morphogens in a concentration-dependent manner to regulate cell fate decisions. This mode of action raises the provocative idea that these same morphogens might also provide a coordinate system for axons to establish the distance to their targets during axon guidance and synapse formation.

Animals↗

Local circuit neurons in both the dentate gyrus and Ammon's horn establish synaptic connections with principal neurons in five day old rats: a morphological basis for inhibition in early development.

Glutamate decarboxylase (GAD)-positive and Golgi impregnated local circuit neurons of the hippocampal formation of five day old rats were examined in light and electron microscopic preparations. The ultrastructural features of these neurons were similar in both the dentate gyrus and CA1 area of Ammon's horn. Somata displayed a perikaryal cytoplasm rich in organelles but lacked organized Nissl bodies. Most nuclei showed intranuclear infoldings of varying degrees but no intranuclear sheets or rods were found. Somata and dendrites were contacted by relatively immature axon terminals that formed mainly symmetric synapses. The axons of local circuit neurons in both the dentate gyrus and Ammon's horn formed symmetric synapses with somata and dendrites of the principal neurons in these regions. Thus, both GAD-positive and Golgi-impregnated terminals of local circuit neurons were observed to form synapses with pyramidal and granule cells. These terminals were usually small and contained relatively few pleomorphic synaptic vesicles. The results show that a circuitry for inhibition is established in the 5 day old dentate gyrus and Ammon's horn, even though the local circuit neurons lack some of the typical adult ultrastructural features at this age.

Animals↗

MuSC, a novel member of the immunoglobulin superfamily, is expressed in neurons of a subset of cranial sensory ganglia in the mouse embryo.

In contrast to the spinal sensory ganglia which reiterate a basic organizational and functional unit, each cranial ganglion mediates a distinct sensory modality and exhibits a characteristic pattern of peripheral and central neuronal connectivity. Molecules responsible for establishment and maintenance of the cranial ganglion-specific networks are not known. Our hamster monoclonal antibody 802C11 strongly stained neurons and their processes of the VIIIth cranial ganglion (hearing and equilibrium), but not of the Vth cranial (somatosensory) or spinal ganglia in the mouse embryo. The cellular staining pattern of positive neurons suggested that the antigen was associated with the cell membrane, and biochemical analyses of the antigen from adult mouse brain showed the antigen to be a glycosylated intrinsic membrane protein of approximately 100 kDa. The antigen was purified, and based on the partial amino acid sequences, its entire cDNA was cloned. A bacterially expressed polypeptide encoded by the cDNA was recognized by the antibody. The deduced amino acid sequence revealed that the antigen belongs to the immunoglobulin superfamily with a significant homology (73.5% identity) to chicken SC1 protein. Chicken SC1 has been shown to be a cell-cell adhesion molecule in vitro with a proposed role in neurite extension of spinal motor neurons. These results suggest that our murine SC1-related protein (MuSC) is involved in the pathfinding and/or fasciculation of specific cranial sensory nerve fibres.

Activated-Leukocyte Cell Adhesion Molecule↗

Reducing contralateral SI activity reveals hindlimb receptive fields in the SI forelimb-stump representation of neonatally amputated rats.

In adult rats that sustained forelimb amputation on the day of birth, >30% of multiunit recording sites in the forelimb-stump representation of primary somatosensory cortex (SI) also respond to cutaneous hindlimb stimulation when cortical GABA(A+B) receptors are blocked (GRB). This study examined whether hindlimb receptive fields could also be revealed in forelimb-stump sites by reducing one known source of excitatory input to SI GABAergic neurons, the contralateral SI cortex. Corpus callosum projection neurons connect homotopic SI regions, making excitatory contacts onto pyramidal cells and interneurons. Thus in addition to providing monosynaptic excitation in SI, callosal fibers can produce disynaptic inhibition through excitatory synapses with inhibitory interneurons. Based on the latter of these connections, we hypothesized that inactivating the contralateral (intact) SI forelimb region would "unmask" normally suppressed hindlimb responses by reducing the activity of SI GABAergic neurons. The SI forelimb-stump representation was first mapped under normal conditions and then during GRB to identify stump/hindlimb responsive sites. After GRB had dissipated, the contralateral (intact) SI forelimb region was mapped and reversibly inactivated with injections of 4% lidocaine, and selected forelimb-stump sites were retested. Contralateral SI inactivation revealed hindlimb responses in approximately 60% of sites that were stump/hindlimb responsive during GRB. These findings indicate that activity in the contralateral SI contributes to the suppression of reorganized hindlimb receptive fields in neonatally amputated rats.

Action Potentials↗

Signal propagation along unidimensional neuronal networks.

Dissociated neurons were cultured on lines of various lengths covered with adhesive material to obtain an experimental model system of linear signal transmission. The neuronal connectivity in the linear culture is characterized, and it is demonstrated that local spiking activity is relayed by synaptic transmission along the line of neurons to develop into a large-scale population burst. Formally, this can be treated as a one-dimensional information channel. Directional propagation of both spontaneous and stimulated bursts along the line, imaged with the calcium indicator Fluo-4, revealed the existence of two different propagation velocities. Initially, a small number of neighboring neurons fire, leading to a slow, small and presumably asynchronous wave of activity. The signal then spontaneously develops to encompass much larger and further populations, and is characterized by fast propagation of high-amplitude activity, which is presumed to be synchronous. These results are well described by an existing theoretical framework for propagation based on an integrate-and-fire model.

Action Potentials↗

Intrinsic primary afferent neurons of the intestine.

After a long period of inconclusive observations, the intrinsic primary afferent neurons of the intestine have been identified. The intestine is thus equipped with two groups of afferent neurons, those with cell bodies in cranial and dorsal root ganglia, and these recently identified afferent neurons with cell bodies in the wall of the intestine. The first, tentative, identification of intrinsic primary afferent neurons was by their morphology, which is type II in the terminology of Dogiel. These are multipolar neurons, with some axons that project to other nerve cells in the intestine and other axons that project to the mucosa. Definitive identification came only recently when action potentials were recorded intracellularly from Dogiel type II neurons in response to chemicals applied to the lumenal surface of the intestine and in response to tension in the muscle. These action potentials persisted after all synaptic transmission was blocked, proving the Dogiel type II neurons to be primary afferent neurons. Less direct evidence indicates that intrinsic primary afferent neurons that respond to mechanical stimulation of the mucosal lining are also Dogiel type II neurons. Electrophysiologically, the Dogiel type II neurons are referred to as AH neurons. They exhibit broad action potentials that are followed by early and late afterhyperpolarizing potentials. The intrinsic primary afferent neurons connect with each other at synapses where they transmit via slow excitatory postsynaptic potentials, that last for tens of seconds. Thus the intrinsic primary afferent neurons form self-reinforcing networks. The slow excitatory postsynaptic potentials counteract the late afterhyperpolarizing potentials, thereby increasing the period during which the cells can fire action potentials at high rates. Intrinsic primary afferent neurons transmit to second order neurons (interneurons and motor neurons) via both slow and fast excitatory postsynaptic potentials. Excitation of the intrinsic primary afferent neurons by lumenal chemicals or mechanical stimulation of the mucosa appears to be indirect, via the release of active compounds from endocrine cells in the epithelium. Stretch-induced activation of the intrinsic primary afferent neurons is at least partly dependent on tension generation in smooth muscle, that is itself sensitive to stretch. The intrinsic primary afferent neurons of the intestine are the only vertebrate primary afferent neurons so far identified with cell bodies in a peripheral organ. They are multipolar and receive synapses on their cell bodies, unlike cranial and spinal primary afferent neurons. They communicate with each other via slow excitatory synaptic potentials in self reinforcing networks and with interneurons and motor neurons via both fast and slow EPSPs.

Animals↗

Anterograde axonal transport, transcytosis, and recycling of neurotrophic factors: the concept of trophic currencies in neural networks.

Traditional views of neurotrophic factor biology held that trophic factors are released from target cells, retrogradely transported along their axons, and rapidly degraded upon arrival in cell bodies. Increasing evidence indicates that several trophic factors such as brain-derived neurotrophic factor (BDNF), fibroblast growth factor (FGF-2), glial cell-line derived neurotrophic factor (GDNF), insulin-like growth factor (IGF-I), and neurotrophin-3 (NT-3), can move anterogradely along axons. They can escape the degradative pathway upon internalization and are recycled for future uses. Internalized ligands can move through intermediary cells by transcytosis, presumably by endocytosis via endosomes to the Golgi system, by trafficking of the factor to dendrites or by sorting into anterograde axonal transport with subsequent release from axon terminals and uptake by second- or third-order target neurons. Such data suggest the existence of multiple "trophic currencies," which may be used over several steps in neural networks to enable nurturing relationships between connected neurons or glial cells, not unlike currency exchanges between trading partners in the world economy. Functions of multistep transfer of trophic material through neural networks may include regulation of neuronal survival, differentiation of phenotypes and dendritic morphology, synapse plasticity, as well as excitatory neurotransmission. The molecular mechanisms of sorting, trafficking, and release of trophic factors from distinct neuronal compartments are important for an understanding of neurotrophism, but they present challenging tasks owing to the low levels of the endogenous factors.

Animals↗

Regeneration of nerve fibres across a colonic anastomosis in the guinea-pig.

Resection and re-anastomosis of the bowel interrupts enteric neuronal pathways. The reestablishment of neuronal connections across a colonic anastomosis was studied using immunohistochemical, retrograde tracing and physiological techniques. In control guinea-pig proximal colon, retrograde labelling with 1,1'-didodecyl-3,3,3,3'-tetramethylindocarbocyanine perchlorate (DiI) revealed that enteric neurons with anally-directed projections are more numerous and have longer axons than orally-projecting neurons. In resected bowel, up to 26 weeks after re-anastomosis, descending neuronal pathways were substantially interrupted. Immunohistochemical labelling of nerve fibres revealed that some enteric nerve fibres did regenerate across narrow regions of the anastomosis, growing preferentially in the oral to anal direction. However, nerve fibres immunoreactive for neurofilament protein triplet were substantially depleted in myenteric ganglia anal to the anastomosis, even after the longest recovery periods, demonstrating that axonal regrowth was limited. This was confirmed in retrograde tracing studies, as no nerve cell bodies oral to an anastomosis were labelled when DiI was placed on myenteric ganglia just anal to the anastomosis. Physiological studies confirmed that regrowth of nerve fibres across the anastomosis occurred and that it was asymmetric, as electrical stimulation led to aboral conduction across the anastomosis more reliably than oral conduction, as measured by circular muscle contraction. After resection and re-anastomosis of the colon, the disruption of neuronal pathways in the enteric nervous system was observed, with limited and preferential re-establishment of aborally-directed long connections.

Anastomosis, Surgical↗

White matter changes in schizophrenia: evidence for myelin-related dysfunction.

Numerous lines of inquiry implicate connectivity as a central abnormality in schizophrenia. Myelination and factors that affect myelination, such as the function of oligodendroglia, are critical processes that could profoundly affect neuronal connectivity, especially given the diffuse distribution of oligodendrocytes and the widespread distribution of brain regions that have been implicated in schizophrenia. Multiple lines of evidence now converge to implicate oligodendroglia and myelin in schizophrenia. Imaging and neurocytochemical evidence, similarities with demyelinating diseases, age-related changes in white matter, myelin-related gene abnormalities, and morphologic abnormalities in the oligodendroglia demonstrated in schizophrenic brains are all examined in light of the hypothesis that oligodendroglial dysfunction and even death, with subsequent abnormalities in myelin maintenance and repair, contribute to the schizophrenic syndrome.

Age Factors↗