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Control of echolocation pulses by neurons of the nucleus ambiguus in the rufous horseshoe bat, Rhinolophus rouxi. I. Single unit recordings in the ventral motor nucleus of the laryngeal nerves in spontaneously vocalizing bats.

The vocal motor control of the larynx was studied with single unit recordings from the efferent motor nucleus (nucleus ambiguus) in the CF-FM-bat Rhinolophus rouxi, spontaneously emitting echolocation sounds. The experiments were performed in a stereotaxic apparatus that allowed differentiation of activities in the recorded nucleus depending on the electrode position (Fig. 1). Echolocation calls and respiration activity were monitored simultaneously, thus it was possible to compare the time course of the motor control activity during respiration with and without concurrent vocalization. Unit discharges were classified as laryngeal motoneuron activity according to their correlation with the time course (onset and end) of echolocation calls and their discharge rate as: Pre-off-tonic, pre-off-phasic, off-pauser, off-tonic, on-chopper, on-tonic, prior-tonic and inhibitory (Fig. 4). The on-chopper and on-tonic discharge patterns were assigned to the motor activity of the lateral cricoarytenoid muscle and the off-pauser and off-tonic discharge patterns to the motor activity of the posterior cricoarytenoid muscle controlling the time course of vocal pulses. Motoneuron activities recorded under the condition of systematically shifted frequencies in the emitted echolocation calls were investigated in Doppler-shift compensating bats responding to electronically simulated echoes. Of all neurons classified as motor control, only units of the pre-off-tonic discharge type (cricothyroid muscle) changed their activity with frequency shifts in the vocalized pulses; they showed a positive linear correlation with the emitted sound frequency (Fig. 6). In addition, single unit activities in strict synchronization to vocalization were recorded, that by their low discharge rate were not valid as motor control, and were considered to represent activities of interneurons or internuclear neurons connecting the nucleus ambiguus with other vocalization- and respiration-centers (Fig. 3c). Electric lesions in the brain stem and iontophoretically applied horseradish peroxidase (HRP) served as references for localization and morphological identification of the recording sites in cell stained brain slices.

Animals↗

Gene expression in learning processes.

It has repeatedly been shown that long-term memory formation involves neuronal gene expression. In this article several different roles for neuronal gene function in a context of learning are considered: maintenance of neural functioning, replenishment of cellular elements that are exhausted in response to massive neuronal stimulation accompanying behavioral training, maintenance of the plastically reorganized neuronal connections, and finally integration of information at the level of transcription factor-promoter interaction. It is strongly advocated that only careful scrutiny of learning-related gene expression phenomena may aid in understanding of the complex learning process.

Animals↗

The microstructure of cortical neuropil before and after decompression in experimental infantile hydrocephalus.

Hydrocephalus is a common clinical disorder and responsible for many pediatric neurological deficits. Relatively little is known about the cellular mechanisms of this disorder and less is known about reconstitution of connectivity following ventricular shunt procedures. In the present studies experimental infantile hydrocephalus produced by kaolin injection was studied in a neonate kitten model. The neuropil of the cerebral cortex was examined in hydrocephalic animals and animals which received a ventriculoperitoneal shunt to reduce ventriculomegaly. The brains were processed for Golgi silver impregnation and electron microscopy to study the detailed dendritic and synaptic architecture. The periventricular region of the hydrocephalic animals exhibited increased extracellular space and signs of neuronal degeneration. Components of the deep neuropil (laminae V-VI) were in disarray and surrounded by edematous extracellular spaces. The superficial neuropil (laminae I-IV), in contrast, appeared intact, but detailed examination showed indications of dendritic degeneration. Shunt procedures successfully restored the cortical mantle to near normal thickness. However, Golgi light microscopy and electron microscopy revealed that dendritic appendage morphology was altered. The results are discussed in regard to development of neuronal connectivity following shunt procedures.

Animals↗

Heparan sulfate proteoglycans in the nervous system: their diverse roles in neurogenesis, axon guidance, and synaptogenesis.

Development of the mammalian nervous system involves generation of neurons from neural stem cells, migration of generated neurons toward genetically determined locations, extension of axons and dendrites, and establishment of neuronal connectivity. Recent progresses revealed diverse role of heparan sulfate proteoglycans in these processes. This article reviews our current knowledge about the functional roles of heparan sulfate proteoglycans in three critical events in mammalian neural development, namely neurogenesis, axon guidance, and synapse development.

Animals↗

Stereotyped pruning of long hippocampal axon branches triggered by retraction inducers of the semaphorin family.

Like naturally occurring neuronal cell death, stereotyped pruning of long axon branches to temporary targets is a widespread regressive phenomenon in the developing mammalian brain that helps sculpt the pattern of neuronal connections. The mechanisms controlling stereotyped pruning are, however, poorly understood. Here, we provide evidence that semaphorins, activating the Plexin-A3 receptor, function as retraction inducers to trigger-stereotyped pruning of specific hippocampal mossy fiber and pyramidal axon branches. Both pruning events are defective in Plexin-A3 mutants, reflecting a cell-autonomous requirement for Plexin-A3. The distribution of mRNAs for Sema3F and Sema3A makes them candidates for triggering the pruning. In vitro, hippocampal neurons respond to semaphorins by retracting axon branches. These results implicate semaphorins as retraction inducers controlling stereotyped pruning in the mammalian brain.

Animals↗

GABA, not glutamate, a primary transmitter driving action potentials in developing hypothalamic neurons.

Neuronal activity is critical for many aspects of brain development. It has often been assumed that the primary excitatory transmitter driving this activity is glutamate. In contrast, we report that during early development, synaptic release of GABA, the primary inhibitory neurotransmitter in the mature brain, is not only excitatory but in addition plays a more robust role than glutamate in generating spike activity in mouse hypothalamic neurons. Based on gramicidin perforated whole cell and extracellular recording, which leave intracellular Cl(-) unperturbed in brain slices and cultures, the GABA(A) receptor antagonist bicuculline induced a dramatic decrease in spike frequency (83% decrease) in developing neurons, three times greater than that generated by glutamate receptor antagonists 2-amino-5-phosphono-pentanoic acid and 6-cyano-7-nitroquinoxalene-2,3-dione. Thus a number of factors related to spike-dependent stabilization of neuronal connections, including Hebbian mechanisms, that are generally applied to glutamate transmission may also participate in stabilization of GABA circuits.

Action Potentials↗

Learning in realistic networks of spiking neurons and spike-driven plastic synapses.

We have used simulations to study the learning dynamics of an autonomous, biologically realistic recurrent network of spiking neurons connected via plastic synapses, subjected to a stream of stimulus-delay trials, in which one of a set of stimuli is presented followed by a delay. Long-term plasticity, produced by the neural activity experienced during training, structures the network and endows it with active (working) memory, i.e. enhanced, selective delay activity for every stimulus in the training set. Short-term plasticity produces transient synaptic depression. Each stimulus used in training excites a selective subset of neurons in the network, and stimuli can share neurons (overlapping stimuli). Long-term plasticity dynamics are driven by presynaptic spikes and coincident postsynaptic depolarization; stability is ensured by a refresh mechanism. In the absence of stimulation, the acquired synaptic structure persists for a very long time. The dependence of long-term plasticity dynamics on the characteristics of the stimulus response (average emission rates, time course and synchronization), and on the single-cell emission statistics (coefficient of variation) is studied. The study clarifies the specific roles of short-term synaptic depression, NMDA receptors, stimulus representation overlaps, selective stimulation of inhibition, and spike asynchrony during stimulation. Patterns of network spiking activity before, during and after training reproduce most of the in vivo physiological observations in the literature.

Action Potentials↗

Peripheral NT3 signaling is required for ETS protein expression and central patterning of proprioceptive sensory afferents.

To study the role of NT3 in directing axonal projections of proprioceptive dorsal root ganglion (DRG) neurons, NT3(-/-) mice were crossed with mice carrying a targeted deletion of the proapoptotic gene Bax. In Bax(-/-)/NT3(-/-) mice, NT3-dependent neurons survived and expressed the proprioceptive neuronal marker parvalbumin. Initial extension and collateralization of proprioceptive axons into the spinal cord occurred normally, but proprioceptive axons extended only as far as the intermediate spinal cord. This projection defect is similar to the defect in mice lacking the ETS transcription factor ER81. Few if any DRG neurons from Bax(-/-)/NT3(-/-) mice expressed ER81 protein. Expression of a NT3 transgene in muscle restored DRG ER81 expression in NT3(-/-) mice. Finally, addition of NT3 to DRG explant cultures resulted in induction of ER81 protein. Our data indicate that NT3 mediates the formation of proprioceptive afferent-motor neuron connections via regulation of ER81.

Afferent Pathways↗

Chemical and electrotonic connections between Aplysia neurons in primary culture.

Dissociated Aplysia neurons will regenerate neurites and form functional connections in primary cell cultures. The specificity of intercellular connectivity in these cultures was investigated by coculturing neurosecretory bag cells with neurons dissociated from the buccal ganglion. It was found that bag-bag and buccal-buccal electrotonic synapses form with high frequency, consistent with previous findings in pure bag and buccal cultures. There is specificity in the formation of these connections, since no bag-buccal electrotonic synapses were observed. Chemical interactions, on the other hand, are present between bag and buccal neurons. In a buccal-bag pair, injection of sufficient depolarizing current into the buccal cell to elicit a train of action potentials leads to a slow hyperpolarizing response in the bag cell. The bag cell hyperpolarization is accompanied by an increase in the cell's input conductance. This connection appears to be unidirectional, produces a voltage shift in the bag cell which is opposite in sign to that in the buccal cell, and is blocked by the removal of Ca2+ from the extracellular medium, indicating that it is mediated by a chemical neurotransmitter. The selective formation of electrotonic synapses in these mixed bag-buccal cultures, together with the presence of chemically mediated interactions, make this system particularly useful for investigating the establishment of intercellular connectivity.

Animals↗

A family of rat CRMP genes is differentially expressed in the nervous system.

Members of the collapsin/semaphorin family play an important role in creating the complex pattern of neuronal connectivity. Inhibition of growth cone motility by chick collapsin is mediated by the intraneuronal protein CRMP-62. We have now isolated four rat sequences that are highly related to chick CRMP-62. All four genes are expressed exclusively in the nervous system and primarily during development. Rat CRMP-2/TOAD-64 is most closely related to chick CRMP-62 and is the most widely expressed CRMP within the nervous system. Rat CRMP-1 and CRMP-4/rUlip are expressed during discrete periods of neuronal development and are not found in the adult nervous system. Rat CRMP-3 has a distinct distribution, being expressed transiently in developing spinal cord and selectively in the postnatal cerebellum. The differential expression of these genes suggests that CRMPs may transduce signals from different semaphorins and that semaphorins may regulate the plasticity of the adult nervous system.

Aging↗

The development of a patchy organization of the rat striatum.

The rat striatum can be divided into patch and matrix compartments. Patches, as marked by high opiate receptor binding, first emerge perinatally from a dense, diffuse field of striatal opiate binding. Our quantitative analysis revealed that the patch compartment formed its peak proportion of the total striatal area at postnatal day 7. After this time, patches occupied a smaller proportion of the striatum, reflecting the fact that the number of patches and mean area per patch reached near adult levels during the first postnatal week, yet the volume of the striatum as a whole continued to increase for several weeks postnatally. Results from transplant and early postnatal lesion experiments suggested that connections between the striatum and other brain areas are important for the formation and/or maintenance of the patch and matrix compartments. Transplants of embryonic striatum to cavities in the cortex of young adult hosts developed diffuse opiate receptor binding but not dopamine receptor binding. Significantly, the opiate receptor binding seen in the transplants was never organized into the dense patches normally seen in the adult striatum. In a few transplants areas of relatively higher opiate receptor binding occurred in areas of relatively low neuronal cell density, as is seen in early normal development, but the dense adult patches never developed. Coronal diencephalic hemisections, but not decortications, in the early postnatal period produced drastic shrinkage of the striatum and, more importantly, a large decrease in opiate receptor patches when expressed as a proportion of total striatal area. Neuronal connections with more caudal brain structures may play a role in the final differentiation and maintenance of the striatal compartments.

Animals↗

Retinofugal and retinopetal connections in the upside-down catfish (synodontis nigriventris).

The retinofugal and retinopetal connections in the upside-down catfish Synodontis nigriventris were studied by use of the horseradish-peroxidase (HRP) techniques, autoradiography, and degeneration-silver methods. An unusual retinal projection to the torus semicircularis as well as projections to the retina from three different sources in the brain are described. After intra-ocular injections of HRP, labeled cells were found in the optic tectum, the dorsomedial optic nucleus and one of the pretectal nuclei. These new findings support the basic hypothesis (i) that neuronal connections are more extensive in primitive brains, and (ii) that the evolutionary development of more complex brains involves the loss of some selected connections.

Animals↗

Cortical network reorganization guided by sensory input features.

Sensory experience alters the functional organization of cortical networks. Previous studies using behavioral training motivated by aversive or rewarding stimuli have demonstrated that cortical plasticity is specific to salient inputs in the sensory environment. Sensory experience associated with electrical activation of the basal forebrain (BasF) generates similar input specific plasticity. By directly engaging plasticity mechanisms and avoiding extensive behavioral training, BasF stimulation makes it possible to efficiently explore how specific sensory features contribute to cortical plasticity. This review summarizes our observations that cortical networks employ a variety of strategies to improve the representation of the sensory environment. Different combinations of receptive-field, temporal, and spectrotemporal plasticity were generated in primary auditory cortex neurons depending on the pitch, modulation rate, and order of sounds paired with BasF stimulation. Simple tones led to map expansion, while modulated tones altered the maximum cortical following rate. Exposure to complex acoustic sequences led to the development of combination-sensitive responses. This remodeling of cortical response characteristics may reflect changes in intrinsic cellular mechanisms, synaptic efficacy, and local neuronal connectivity. The intricate relationship between the pattern of sensory activation and cortical plasticity suggests that network-level rules alter the functional organization of the cortex to generate the most behaviorally useful representation of the sensory environment.

Acetylcholine↗

Facial nerve parasympathetic preganglionic afferents to the accessory otic ganglia by way of the chorda tympani nerve in the cat.

The distribution of accessory otic ganglia and connections between the ganglia and the chorda tympani nerve were investigated in the cat in order to determine the parasympathetic preganglionic facial nerve afferents to the otic ganglia using whole mount acetylthiocholinesterase (WATChE) histochemistry. The otic ganglia consist of a single main prominent ganglion and many small accessory ganglia lying on a plexus around the origins of the branches of the mandibular nerve and near the junction of the chorda tympani nerve and lingual nerve. In cell analysis of Nissl-stained preparations, the neurons composing the accessory otic ganglia were morphologically similar to the main otic ganglion neurons. Connecting branches from the chorda tympani nerve to the peripherally located accessory otic ganglia were found and they were not stained by WATChE histochemistry. WATChE-positive connecting branches from the ganglia to the inferior alveolar, lingual, and mylohyoid nerves were also found in the same preparations. The WATChE histochemistry on various autonomic nervous tissues revealed that autonomic postganglionic nerve fibers are selectively stained darkly and that preganglionic fibers remain unstained. Therefore, it is considered that the WATChE-negative connections from the chorda tympani nerve consist chiefly of autonomic preganglionic fibers, whereas the WATChE-positive connections to the branches of the mandibular nerve are mainly postganglionic fibers. This suggests that some of the facial nerve parasympathetic preganglionic fibers in the chorda tympani nerve are mediated in the accessory otic ganglia and then join the branches of the mandibular nerve to supply the target mandibular tissues.

Acetylcholinesterase↗

The septointerpeduncular projection in the rat: tracing with the carbocyanine dye Dil.

The fluorescent carbocyanine dye Dil has been used to retrogradely label the neuronal connections between the forebrain septal area and the interpeduncular nucleus. Previous works based on retrograde horseradish peroxidase transport have identified that only the diagonal band nucleus is a source of the septointerpeduncular projections, but anterograde tracing with labeled amino acids and selective lesions with colchicine have shown that also the posterior septal nuclei project to the interpeduncular nucleus. In the present study, the retrograde labeling in septal nuclei after placing the carbocyanine Dil in the interpeduncular nucleus resulted in the fluorescent labeling of numerous neurons of the diagonal band nucleus. Our results, in addition, showed the labeling of some scattered neurons in the ventral portion of the triangular nucleus of the septal area and in the septofimbrial nucleus, confirming the presence of a previously controversial septointerpeduncular projection.

Animals↗

Alzheimer's disease as a loss of differentiation control in a subset of neurons that retain immature features in the adult brain.

Degeneration in AD primarily occurs in a subset of neurons that in the adult brain retain a high degree of structural plasticity and in these neurons is associated with the activation of mitogenic pathways and a cell cycle re-entry. Brain areas affected by AD pathology are those structures involved in the regulation of "higher brain functions" that become increasingly predominant as the evolutionary process of encephalization progresses, such as hippocampus, neocortical association areas and the cholinergic basal forebrain neurons. The functions these areas subserve such as learning, memory, perception, self-awareness, and consciousness require a life-long re-fitting of synaptic contacts that allows for the acquisition of new epigenetic information. This adaptive reorganization of neuronal connectivity in the mature brain is based upon the strengthening of existing synapses, the formation of new synapses and the destabilization of previously established synaptic contacts. With the increasing need during evolution to organize brain structures of increasing complexity, these processes of dynamic stabilization and de-stabilization become more and more important but might also provide the basis for an increasing rate of failure. A hypothesis is proposed that it is the 'labile state of differentiation' (G0-arrest) of a subset of neurons in the adult brain that allows for ongoing morphoregulatory processes after development is completed but at the same time renders these neurons particularly vulnerable. The delicate balance between G0-arrest and G1-entry might be prone to a variety of potential disturbances during the lifetime of an individual. Morphodysregulation in AD, accompanied by an activation of intracellular mitogenic signaling might, thus, be a slowly progressing dysfunction that eventually overrides the differentiation control and results in dedifferentiation, a condition in conflict with the otherwise 'mature' background of the nervous system. Cell-cycle and differentiation control might thus provide the link between structural brain self-organization and neurodegeneration that both are unique to human.

Adult↗

Gene transfer: manipulating and monitoring function in cells and tissues.

1. The ectopic expression of genes has proven to be an extremely valuable tool for biologists. The most widely used systems involve electrically or chemically mediated transfer of genes to immortalized cell lines and, at the other end of the spectrum, transgenic animal models. As would be expected, there are compromises to be made when using either of these broad approaches. Immortalized cell lines have limited "physiological relevance" and transgenic approaches are costly and out of the reach of many laboratories. There is also significant time required for the de novo generation of a transgenic animal. 2. As a viable alternative to these approaches, we describe the use of recombinant adenovirus and Sindbis virus to deliver genes to cells and tissues. 3. We exemplify this approach with studies from our laboratories: (i) an investigation of Ca2+ handling deficits in cardiac myocytes of hypertrophied hearts using infection with recombinant adenovirus encoding either green fluorescent protein (GFP) or the sarcoplasmic/endoplasmic reticulum calcium-ATPase (Serca2a); (ii) a study of the mechanism of macrophage/microglial migration by infection of embryonic phagocytes with a GFP-encoding virus and coculture with brain slices to then track the movement of labelled cells; and (iii) we are also exploiting the natural tropism of the Sindbis virus to label neurons in hippocampal brain slices in culture to resolve high-resolution structure and to map neuronal connectivity. 4. Further development of these approaches should open new avenues of investigation for the study of physiology in a range of cells and tissues.

Adenoviridae↗

The telencephalon of tetrapods in evolution.

Numerous scientists have sought a homologue of mammalian isocortex in sauropsids (reptiles and birds) and a homologue of sauropsid dorsal ventricular ridge in mammals. Although some of the proposed theories were enormously influential, alternative theories continued to coexist, primarily because the striking differences in pallial organization between adult mammals, sauropsids, and amphibians enabled different authors to enlist different subsets of similarity data in support of different hypotheses of putative homology. A phylogenetic analysis based on parsimony cannot discriminate between such alternative hypotheses of putative homology, because sauropsids and mammals are sister groups. One solution to this dilemma is to include embryological patterns of telencephalic organization in the comparative analysis. Because early developmental stages in different taxa tend to resemble each other more than the adults do, the embryological data may reveal intermediate patterns of organization that provide unambiguous support for a single hypothesis of putative homology. The validity of this putative homology may then be supported by means of a phylogenetic analysis based on parsimony. A comparative analysis of pallial organization that includes embryological data suggests the following set of homologies. The lateral cortex in reptiles is homologous to the piriform cortex in birds and mammals. The anterior dorsal ventricular ridge in reptiles is probably homologous to the neostriatum and ventral hyperstriatum in birds and to the endopiriform nucleus in mammals. The posterior dorsal ventricular ridge in reptiles is most likely homologous to the archistriatum in birds and to the pallial amygdala in mammals. The pallial thickening in reptiles is probably homologous to the dorsal and intercalated portions of the hyperstriatum in birds and to the claustrum proper in mammals. Finally, the dorsal cortex in reptiles is probably homologous to the accessory hyperstriatum and parahippocampal area in birds and to the isocortex in mammals. These hypotheses of homology imply relatively minor evolutionary changes in development but major changes in neuronal connections. Most significantly, they imply the independent elaboration of thalamic sensory projections to derivatives of the lateral and dorsal pallia in sauropsids and mammals, respectively. They also imply the independent evolution of lamination in the pallium of birds and mammals.

Animals↗