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The development of projections and connections from transplanted locust sensory neurons.

Neurons innervating wind-sensitive hairs on the locust head form characteristic projections and connections within the CNS. These depend on intrinsic properties of the epidermis from which the hair and its neuron are formed (Anderson & Bacon, 1979; Bacon & Anderson, 1984). To investigate further these intrinsic properties and also extrinsic factors involved in guiding axon growth and determining synaptic connectivity, pieces of epidermis from the head were transplanted to the posterior head, prothorax, or mesothorax. Thus wind-sensitive neurons developing from the grafts were caused to grow into foreign parts of the CNS. The neuronal projections from the graft hairs were examined by filling the axons with cobalt, and their connectivity with an identified interneuron, the Tritocerebral Commissure Giant, was examined by recording electrophysiologically the activity of the interneuron during stimulation of the graft hairs. The results show that the neuronal projections are confined to one tract, the median ventral tract, and to one arborization area, the ventral association centre, in all ganglia; in all ganglia, neurons from different epidermal regions preserve their location-specific properties of forming ipsilateral or additional contralateral projections; the extent of their projection in the CNS is not interpretable in terms of intrinsic instructions only; in foreign ganglia, they fail to form connections with their normal target interneuron.

Animals↗

Neuronal organization and cell interactions of the cat stellate ganglion.

The functional structure of the cat stellate ganglion (SG) and, in particular, its extra- and intraganglionic connections and neuronal organization, were investigated using histochemical, immunohistochemical, morphological and histological methods. Retrograde axonal transport of horseradish peroxidase was used to determine most of the extraganglionic interactions. Of the targets tested, the most extensive efferent connections of the SG were with the stemocleidomastoid muscle, trachea, esophagus and heart. Neurons of the SG also send a small number of postganglionic efferents to the thyroid and stomach. Furthermore, ganglion cells send axons to the spinal ganglia. Several afferent connections of the SG were determined. Sympathetic preganglionic neurons of segments C8-T10 of the spinal cord, sensory neurons of C8-T9 spinal ganglia, intramural ganglia of the thoracic viscera and the reticular formation of the medulla oblongata send their axons to the SG. Intraganglionic interactions of intemeurons with principal ganglionic cells were assumed to occur, based on the presence of interneurons immunoreactive to GABA and substance P. GABA- and substance P-immunoreactive fibers located around a small number of postganglionic neurons were also identified. Morphological study revealed asymmetry between the left and right ganglia: the right ganglion is larger than the left and contains more cells. This asymmetry was also reflected in basic structural parameters of neurons, such as average neuronal area and average diameter of cell somata. The present data has been used to develop a scheme for the basic inter- and intraneuronal connections of the cat SG. This ganglion is a true nervous center, with postganglionic neurons, some of which might be performing sensory functions, and interneurons. The ganglion is connected not only with the spinal cord and spinal ganglia, but also with neurons of the intramural ganglia and, by direct links, with efferent neurons of the medulla oblongata. Thus, the SG may play an essential role in viscera-visceral reflexes.

Afferent Pathways↗

Synaptic connections of neuropeptide Y (NPY) immunoreactive neurons in the hilar area of the rat hippocampus.

Synaptic connections and fine structural characteristics of neuropeptide Y-immunoreactive (NPY-i) neurons in the fascia dentata were studied using an antiserum against NPY. Normal and colchicine pretreated rats were examined to study the synaptic connections of NPY-i neurons in the normal fascia dentata. The perforant pathway and fimbria fornix were transected to label afferent fibers to NPY-positive cells. Horseradish peroxidase conjugated with wheat germ agglutinin (HRP-WGA) was injected into the contralateral hippocampus to study commissural projections of hippocampal NPY-i neurons, and to search for NPY-i synaptic contacts on immunonegative commissural cells. Since earlier reports have shown that at least half of the NPY-i neurons also contain somatostatin (SS), the distribution of NPY-i neurons in the hilar area was determined and compared with that of SS-i neurons. Four types of dentate NPY-i neurons were distinguished: Type 1: large multipolar cells in the deep hilus (9%). Type 2: medium-sized multipolar and fusiform hilar neurons with dendrites occasionally reaching the outer molecular layer (64%). Type 3: pyramidal shaped cells in the granule cell layer with long apical dendrites reaching the outer molecular layer (20%). Type 4: small multipolar NPY-i cells located in the molecular layer (7%). Our results indicate two overlapping but not identical cell populations of NPY-i and SS-i neurons. Light and electron microscopic analysis of the normal fascia dentata demonstrated that the majority of NPY-i terminals are located in the outer molecular layer of the dentate gyrus, where they establish symmetric synaptic contacts on dendritic shafts and occasionally on spines of granule cells. A moderate number of NPY-i synapses were also found on dendrites in the inner molecular layer and on the cell body of granule cells. Numerous symmetric NPY-i synapses were found on dendrites and somata of neurons in the hilar area. Some NPY-i dendrites in the hilar area received mossy axon collateral input. After transection of the perforant pathway degenerated axon terminals could be found in synaptic contact with NPY-i dendrites in the outer molecular layer. Commissurotomy revealed direct commissural input to NPY-i dendrites in the inner molecular layer and in the hilus. After injection of HRP-WGA into the contralateral hippocampus 2% of hilar NPY-i neurons were retrogradely labeled and symmetric NPY-i synapses were found on the cell bodies and dendrites of unstained HRP-WGA labeled neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Direct projection from the dorsal hypothalamic area to the nucleus raphe pallidus: a study using anterograde transport with Phaseolus vulgaris leucoagglutinin in the rat.

A hypothalamic projection to the nucleus raphe pallidus of the medulla was examined using the anterograde tracing technique based on Phaseolus vulgaris leucoagglutinin (PHA-L) in the rat. After the iontophoretic application of PHA-L to the dorsal hypothalamic area, labeled fibers that finally ended in the nucleus raphe pallidus were observed descending through the most medial part of the ventral tegmental area and the nucleus reticularis tegmenti pointis to reach the medial aspect of the pyramid. Many varicose fibers forming a loose plexus were observed in the nucleus raphe pallidus, especially ventrally. The ventral surface of the pyramid and the most ventral region of the nucleus reticularis paragigantocellularis lateralis (PGCL) contained labeled varicose fibers. At the electron microscopic level, the labeled profiles in the nucleus raphe pallidus were small-sized unmyelinated axons and axon terminals. Labeled axon terminals containing spherical synaptic vesicles formed synapses on spine-like protrusions or small-sized dendritic shafts. These results strongly indicate that neurons in the dorsal hypothalamic area have a direct connection with neurons in the nucleus raphe pallidus and the ventral part of the PGCL. The possible involvement of this pathway in cardiovascular regulation was discussed.

Animals↗

[A new method of analysis of impulse activity in brain neurons].

Investigation of impulse activity of hypothalamic neurons is of a great importance in attempt to connect the neuronal function with its activity. We have proposed an original method of analysis of background and temperature-induced impulse activity of thermosensitive neurons from frontal hypothalamus. In acute experiments on cats under ketamine anastasia neuronal activity was measured by glass microelectrode. Measurements of background neuronal activity was performed during 30 s, and post-stimulus activity was measured during 60 s. Computer program calculated the mean value of background activity. These results were used to create bar graph in the range +/- 100% with time interval 10%. It is suggested that the results of analysis of background activity of thermosensitive hypothalamic neurons allow interpret the characteristics of functions of hypothalamic neurons.

Animals↗

[The multiplicity of neurotransmitters: the functional significance].

The generally accepted synaptic concept is an idealization based on two major assumptions essential for a system of neurone to be orderly organized: firstly, specificity of anatomical connections between neurones is assumed, and, secondly, it is postulated that the signal is transmitted via isolated compartment of extracellular space, the synaptic cleft. In the framework of this conceptual model, systems can be built using a single sort of signal molecules, and knowledge on diversity of neurotransmitters remains unconceptualized. To provide that the signal is properly delivered to a due address, neurones can however equally utilize the chemical mode of addressing. According to an alternative conceptual model proposed by the author, orderliness of a neuronal system is based on plurality of signal molecules. To describe the idealized elementary unit of heterochemical integration, the term "Heteron" is introduced. Heteron is defined as the network of individually specific neurones differing in their respective transmitters. It is postulated that (i) the transmitter situation is changed evenly throughout the heteron and (ii) responses of sensitive targets to a specific transmitter situation are composed into a well co-ordinated whole. The functional repertory of a heteron containing n neurones (i.e. n transmitters) will thus include at least n integrated states. The conceptual synapse corresponds to a theoretical limit of the heteron (when n = 1). It is suggested that primitive, simpler nervous systems roughly correspond to a single heteron while evolved brains and cords include multiple units of heterochemical integration.

Animals↗

Synaptic connections and small circuits involving excitatory and inhibitory neurons in layers 2-5 of adult rat and cat neocortex: triple intracellular recordings and biocytin labelling in vitro.

Dual and triple intracellular recordings with biocytin labelling in slices of adult neocortex explored small circuits of synaptically connected neurons. 679 paired recordings in rat and 319 in cat yielded 135 and 42 excitatory postsynaptic potentials (EPSPs) and 37 and 26 inhibitory postsynaptic potentials (IPSPs), respectively. Patterns of connectivity and synaptic properties were similar in the two species, although differences of scale and in the range of morphologies were observed. Excitatory 'forward' projections from layer 4 to 3, like those from layer 3 to 5, targeted pyramidal cells and a small proportion of interneurons, while excitatory 'back' projections from layer 3 to 4 selected interneurons, including parvalbumin immuno-positive basket cells. Layer 4 interneurons that inhibited layer 3 pyramidal cells included both basket cells and dendrite-targeting cells. Large interneurons, resembling cells previously described as large basket cells, in layers 4 and 3 (cat), with long myelinated horizontal axon collaterals received frequent excitatory inputs from both layers. A very high rate of connectivity was observed between pairs of interneurons, often with quite different morphologies, and the resultant IPSPs, like the EPSPs recorded in interneurons, were brief compared with those recorded in pyramidal and spiny stellate cells.

Animals↗

Selective stabilization and synaptic specificity: a new cell-biological model.

How are appropriate connections between neurons sorted from the overwhelming surplus of potential, yet inappropriate, connections? Despite the apparently improbable nature of the process, brains wire themselves with a high degree of reproducibility that has been conserved across evolutionary history. Here, we outline a viable cell-biological model for generating synaptic specificity that features selection of nascent synapses based on adhesion and recognition. This process uses the highly dynamic and stochastic nature of intracellular trafficking to generate reproducible patterns of synaptic connectivity.

Animals↗

[Interrelationship between differently spaced neurons in the auditory and motor zones of the cerebral cortex in cats].

Multineuronal activity was studied in unrestrained cats with bundles of seven electrodes (tips dia. 50 mcm) chronically implanted in the auditory and motor cortex. Spikes of the highest and lowest amplitude were isolated from each neurogram by an amplitude discriminator. Functional connections between neurones located under different electrodes at a distance of up 540 mcm were defined by statistical dependence of the isolated spike series. In the auditory cortex, the distance at which it was possible to detect connections between high amplitude neurones (H-H) did not exceed 450 mcm, while between low amplitude neurones (L-L) it was 270 mcm. In the motor cortex, dependent relations for similar pairs of neurones (H-H) and (L-L) were found at all the studied distances up to 540 mcm. For mixed neuronal pairs (H-L and L-H) functional connections at different distances were differently expressed at various distances.

Animals↗

Enhanced excitatory synaptic connectivity in layer v pyramidal neurons of chronically injured epileptogenic neocortex in rats.

Formation of new recurrent excitatory circuits after brain injuries has been hypothesized as a major factor contributing to epileptogenesis. Increases in total axonal length and the density of synaptic boutons are present in layer V pyramidal neurons of chronic partial isolations of rat neocortex, a model of posttraumatic epileptogenesis. To explore the functional consequences of these changes, we used laser-scanning photostimulation combined with whole-cell patch-clamp recording from neurons in layer V of somatosensory cortex to map changes in excitatory synaptic connectivity after injury. Coronal slices were submerged in artificial CSF (23 degrees C) containing 100 microM caged glutamate, APV (2-amino-5-phosphonovaleric acid), and high divalent cation concentration to block polysynaptic responses. Focal uncaging of glutamate, accomplished by switching a pulsed UV laser to give a 200-400 micros light stimulus, evoked single- or multiple-component composite EPSCs. In neurons of the partially isolated cortex, there were significant increases in the fraction of uncaging sites from which EPSCs could be evoked ("hot spots") and a decrease in the mean amplitude of individual elements in the composite EPSC. When plotted along the cortical depth, the changes in EPSCs took place mainly between 150 and 200 microm above and below the somata, suggesting a specific enhancement of recurrent excitatory connectivity among layer V pyramidal neurons of the undercut neocortex. These changes may shift the balance within cortical circuits toward increased synaptic excitation and contribute to epileptogenesis.

2-Amino-5-phosphonovalerate↗

Major role for neuronal death during brain development: refinement of topographical connections.

The precision of the topographic projection from the isthmo-optic nucleus (ION) to the retina has been examined in chicken embryos and chicks by the retrograde transport of a fluorescent carbocyanine dye from restricted retinal sites. At all ages, the labeled neurons are most numerous in the topographically appropriate part of the ION, but in younger embryos up to 49% of them are found outside this region. The distribution of these "aberrantly" projecting neurons is variable, but they generally occur throughout the entire ION. They all die during the ION's period of neuronal death, accounting for most of the 60% cell loss that then occurs. We therefore suggest that a major role of neuronal death during brain development is to reduce the imprecision of neuronal connections.

Animals↗

Functional organization of local neural networks in the cat neocortex. Relationship to the level of food motivation.

Implanted semimicroelectrodes were used to record multineuron activity--spike discharges from groups of close-lying neurons--in the deep layers of the frontal and motor cortex in conscious cats with different levels of food motivation. Spike activity from 4-7 neurons was extracted from multineuron activity, and interneuron interactions were studied by cross-correlation analysis. Neurons in local networks were divided into two subgroups: neurons with high-amplitude spikes with a predominance of output (divergent) connections, and neurons with low-amplitude spikes and a predominance of input (convergent) connections. Food deprivation lasting 24 h affected mainly the nature of interneuron interactions in the range of late cross-correlational connections (with delays of 2-100 msec).

Animals↗

[The temporal organization of the functional connection of the motor cortex neurons in the cat].

Conditioned food-procuring response to time (2 minutes interval) was elaborated in cats, multiunit activity of the motor cortex being recorded. On the basis of single spike trains discriminated from the multiunit activity the cross-correlation histograms were built and the spikes composing their peaks were analysed in real time. This secondary analysis of the histograms allowed to ascertain the dynamics of functional connections between the neurons during the phase of active waiting according to the distribution of coincident impulses. A concentration of coincident impulses of simultaneously recorded cells was observed in different moment of time. In some neuronal pairs the concentration of coincident impulses was revealed to the end of the conditioned interval. The data obtained are considered as a manifestation of the conditioned reaction at the level of neuronal interaction.

Animals↗

Peptidergic neurotransmitters in the endocrine hypothalamus.

More than 20 neuropeptides have been localized in the endocrine hypothalamus. They may exert a neurohormonal effect on the pituitary or innervate other neurons (intranuclear, intrahypothalamic or extrahypothalamic) and act as neurotransmitters. Many of the hypothalamic neuropeptides are synthesized as inactive precursors that are activated by proteolysis during axonal transport from the cell body to the synapse. Studies in which the paraventricular nuclei were bilaterally destroyed have shown that the neuroendocrine cells in the hypothalamus show functional plasticity and cells that do not usually make detectable quantities of a particular neuropeptide may be activated to do so. Within the hypothalamic nuclei are dense networks of synaptic connections among neurons synthesizing the same or different neuropeptides. These local circuits may coordinate the activities of peptidergic neurons in a hypothalamic nucleus. Hypothalamic neurons project axons to the median eminence-pituitary stalk and the posterior pituitary, also to nuclei within the hypothalamus and to extrahypothalamic areas such as the lower brainstem. Peptidergic neurons in the hypothalamus can have combined neurohormonal and neurotransmitter activities mediated by axon terminals on portal capillaries and other hypothalamic nuclei. Double labelling immunohistochemistry has been used to demonstrate reciprocal connections between peptidergic neurons in the hypothalamus, such as those synthesizing growth hormone-releasing hormone and somatostatin.

Animals↗

Synaptic connections, axonal and dendritic patterns of neurons immunoreactive for cholecystokinin in the visual cortex of the cat.

A subpopulation of gamma-aminobutyric acid (GABA) containing neurons was reported to contain cholecystokinin-immunoreactive material in the visual cortex of cat [Somogyi et al., J. Neurosci. (1984) 4, 2590-2603]. In the present study pre-embedding immunocytochemistry was used to identify which of the several types of presumed GABAergic nonpyramidal cells in areas 17 and 18 contain cholecystokinin immunoreactivity. Most of the cholecystokinin-immunoreactive somata were found in layers II-III, they were less frequent in layers I and VI, and relatively rare in layers IV and V. The distribution and density of the axon terminals resembled that of the cell bodies. Two well defined types of cholecystokinin-immunoreactive neuron were distinguished: (1) double bouquet cells in layers II-III with vertically projecting axons, and (2) small basket cells with local axons either restricted to layers II-III, or descending to layer V. Additional cholecystokinin-positive cells showed features of bitufted or multipolar neurons in layers II-VI and horizontal cells in layer I, but these cells could be defined less well due to partial staining. Cholecystokinin-immunoreactive dendrites were found to run horizontally in layer I for several hundred micrometers. Some of the cholecystokinin-immunoreactive cells in layer VI had very long dendrites ascending radially up to layer III, as did their axons. A few cholecystokinin-immunoreactive cells appeared to have two axons and this was confirmed by electron microscopy. All cholecystokinin-immunoreactive neurons and terminals were separated from the basal lamina of blood vessels by glial endfeet. Random samples of boutons from each layer as well as identified terminals traced to their origin from local neurons were examined in the electron microscope. All of the boutons established symmetrical (type II) synaptic contacts with dendritic shafts, spines or somata. Quantitative electron microscopy of the postsynaptic targets of double bouquet cells and small basket cells demonstrated clear differences between these two types of neuron; basket cells having a higher proportion of their terminals in synaptic contact with somata. The findings that several distinct types of cortical neurons, as defined by their synaptic connections, contain cholecystokinin-immunoreactive material and that identified axons of all examined neurons form type II synaptic contacts suggests that the majority, if not all cholecystokinin-positive boutons forming type II contacts originate from local cortical cells. The distribution of targets postsynaptic to cholecystokinin-positive neurons is compared to those of cells labelled by other methods.

Animals↗

Effects of static and dynamic disorder on the performance of neural automata.

We report on both analytical and numerical results concerning stochastic Hopfield-like neural automata exhibiting the following (biologically inspired) features: (1) Neurons and synapses evolve in time as in contact with respective baths at different temperatures; (2) the connectivity between neurons may be tuned from full connection to high random dilution, or to the case of networks with the small-world property and/or scale-free architecture; and (3) there is synaptic kinetics simulating repeated scanning of the stored patterns. Although these features may apparently result in additional disorder, the model exhibits, for a wide range of parameter values, an extraordinary computational performance, and some of the qualitative behaviors observed in natural systems. In particular, we illustrate here very efficient and robust associative memory, and jumping between pattern attractors.

Models, Neurological↗

Analysis of neuronal networks: a review of techniques for labeling axonal projections.

In order to analyze connections between neurons in the vertebrate central nervous system, methods have been developed to label a given population of axons of known origin so that they can be differentiated from other, non-labeled structures. Three such methods are reviewed here: experimentally induced orthograde (Wallerian) degeneration, axon transport of radioactive proteins demonstrated by autoradiography, and axon transport of macromolecules that can be reacted histochemically to yield a visible reaction product. Each of the methods has particular strengths and weaknesses. Degeneration methods may differentiate between different functional classes of axons which have different fiber diameters. However, degeneration distorts the morphology of axon terminals, making them more difficult to interpret, and degenerating terminals may be removed rapidly by phagocytosis. Autoradiography of radioactive terminals preserves normal fine structure, but the necessary exposure times extend the method by weeks or months, and care must be exercised to distinguish labeled axons from other structures exhibiting background or transneuronal radioactivity. Histochemical methods, such as those used to demonstrate horseradish peroxidase conjugated to wheat germ lectin (WGA-HRP), are sensitive and rapid, but the injection site must be carefully characterized, and the presence of transneuronal label may make interpretation of the results difficult. Experimental methods of axonal labeling have been invaluable in studying neuronal networks. Each of the methods described here may be of particular value, given the nature of the system to be analyzed.

Animals↗

Tooth pulp primary neurons: cell size analysis, central connection, and carbonic anhydrase activity.

Cell bodies and central terminals of trigeminal primary afferent neurons innervating the mandibular molar and incisor tooth pulps were labeled by injecting various neuroanatomical tracers into these tooth pulps. The cell bodies of major constituents of primary neurons innervating the tooth pulp, cornea, and cutaneous branch of the mylohyoid nerve were large (> or = 500 microns2, 65%), medium (300-400 microns2, 35%), and small (100-200 microns2, 34%), respectively. Those innervating the tooth pulp (tooth pulp cells) had the Nissl pattern characteristic of large light cells of the A type, while those innervating the cornea were small dark cells of the B type. Thirty percent of the tooth pulp cells exhibited histochemically demonstrable carbonic anhydrase activity. The transganglionic transport of HRP-WGA indicated marked concentration of central terminals of the tooth pulp primaries in the rostral subdivisions of the brain stem sensory trigeminal nuclear complex. In contrast, central terminals of the corneal primaries were concentrated in the medullary dorsal horn.

Animals↗