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Protein-synthesizing machinery in the axon compartment.

Contrary to the prevailing view that the axon lacks the capacity to synthesize proteins, a substantial body of evidence points to the existence of a metabolically active endogenous translational machinery. The machinery appears to be largely localized in the cortical zone of the axon, where, in vertebrate axons, it is distributed longitudinally as intermittent, discrete domains, called periaxoplasmic plaques. Studies, based on translation assays and probes of RNA transcripts in axon models such as the squid giant axon and selected vertebrate axons, provide evidence of locally synthesized proteins, most of which appear to be constituents of the slow axoplasmic transport rate groups. Metabolic and molecular biological findings are consistent with the view that the synthesis of proteins undergoing local turnover in the axonal compartment of macroneurons depends on the activity of an endogenous translational machinery. The documented presence of a metabolically active machinery in presynaptic terminals of squid photoreceptor neurons is also described. Finally, potential sources of axoplasmic RNAs comprising the machinery, which may include the ensheathing cell of the axon, as well as the cognate cell body, are also discussed.

Animals↗

Taurine release modified by nitric oxide-generating compounds in the developing and adult mouse hippocampus.

The effects of the nitric oxide-generating compounds hydroxylamine, sodium nitroprusside and S-nitroso-N-acetylpenicillamine, and the nitric oxide synthase inhibitors nitroarginine and 7-nitroindazole on taurine release from hippocampal slices from adult (three-month-old) and developing (seven-day-old) mice were characterized using a superfusion system. The basal release of [3H]taurine was enhanced when the nitric oxide donors were added at the beginning of superfusion, more markedly in the adult than in the immature hippocampus. The effect of hydroxylamine was clearly concentration-dependent. Hydroxylamine also markedly enhanced the release of endogenous taurine. The K+-stimulated (50 mM) release of taurine was generally inhibited by the nitric oxide-generating compounds in both age groups. Nitric oxide is thus able to act directly at presynaptic terminals, modulating taurine release as a retrograde messenger. The N-methyl-D-aspartate-evoked taurine release was reduced by the nitric oxide donors, particularly by sodium nitroprusside, in the adult hippocampus, while hydroxylamine and S-nitroso-N-acetylpenicillamine markedly potentiated the release in developing mice. In the immature hippocampus the hydroxylamine-enhanced taurine release seems to involve a Ca2+-independent, Na+-dependent and carrier-mediated process while in adult mice only a part of the hydroxylamine-enhanced release is mediated by the same mechanism. The results show that nitric oxide-generating compounds modify the basal, K+- and N-methyl-D-aspartate-evoked releases of taurine in both adult and immature hippocampus. The enhanced N-methyl-D-aspartate receptor-evoked release may be an important mechanism protecting the immature brain against excitotoxicity.

Aging↗

High level of adenosine A1 receptor-like immunoreactivity in the CA2/CA3a region of the adult rat hippocampus.

We describe the immunocytochemical distribution of adenosine A1 receptors in the rat hippocampus. Adenosine A1 receptor-like immunoreactivity was seen on the cell soma and dendrites of pyramidal cells and the cell soma and proximal part of dendrites of granule cells, but not on glial cells. Developmentally, adenosine A1 receptor-like immunoreactivity was diffuse on postnatal day 7 and increased in intensity in individual cells by day 21. In the CA2/CA3a region, the adult pattern of A1 receptor distribution was established by day 28. In the adult rat hippocampus, rostrocaudal inspection revealed that immunoreactivity in CA2/CA3a was greatest. Confocal microscopy revealed differences in the staining patterns for the adenosine A receptor and synaptophysin, a marker of presynaptic terminals. This result suggests that the adenosine A1 receptor might have postsynaptic physiological functions. Double-labeling of adenosine A1 receptors and anterogradely-labeled fibers from the supramammillary nucleus showed that the fibers from the supramammillary nucleus terminate directly on the cell soma of the A1 receptor-immunopositive neurons in CA2/CA3a and the dentate gyrus. These results indicate that the adenosine A 1 receptor in CA2/CA3a and the dentate gyrus are in a position to regulate hippocampal theta activity and that resultant strong synaptic depression in CA2/CA3a could play a role in regulating the intrinsic signal flow between CA3 and CA1.

Adenosine↗

Autoradiographic comparison of [3H](-)nicotine, [3H]cytisine and [3H]epibatidine binding in relation to vesicular acetylcholine transport sites in the temporal cortex in Alzheimer's disease.

The laminar binding distribution of three nicotinic receptor agonists, [3H](-)nicotine, [3H]cytisine, and [3H]epibatidine, and their relation to the [3H]vesamicol binding, which is known to represent the vesicular acetylcholine transport sites, was performed employing in vitro autoradiography on the medial temporal cortex (Brodmann area 21). Autopsied brain tissue from nine Alzheimer patients and seven age-matched controls were used. The binding pattern of the three nicotinic ligands in the normal cortex was in general similar, showing binding maxima in the cortical layers I, III and V. The binding of [3H](-)nicotine, [3H]cytisine, and [3H]epibatidine was lower in the older controls and more uniform throughout the layers as compared with younger controls. There was a significant age-related decrease in the binding of the three nicotinic ligands within the controls (age range: 58 to 89 years; P[3H](-)nicotine = 0.002, P[3H]epibatidine = 0.010, P[3H]cytisine = 0.037). In the older controls, the [3H]epibatidine binding was much decreased as compared with that of [3H](-)nicotine and [3H]cytisine. This may indicate a higher selectivity of [3H]epibatidine for a nicotinic receptor subtype that is particularly affected by aging. The laminar binding pattern of [3H]vesamicol showed one maximum in the outer cortical layers II/III. The [3H]vesamicol binding did not change with aging. The binding of all ligands was significantly decreased in all layers of the temporal cortex in Alzheimer's disease, but the [3H]vesamicol binding decreased only half as much as the nicotinic receptors. Also, choline acetyltransferase activity was percentually more reduced than [3H]vesamicol binding in Alzheimer's disease. The cortical laminar binding pattern of all 3H-ligands was largely absent in the Alzheimer's disease cases. The less severe loss of vesicular acetylcholine transport sites as compared with the loss of the nicotinic receptors and choline acetyltransferase activity may suggest that vesamicol binding sites might be more preserved in presynaptic terminals still existing and thereby expressing compensatory capacity to maintain cholinergic activity.

Acetylcholine↗

Kindling induces a long-term enhancement in the density of N-type calcium channels in the rat hippocampus.

How seizures arise and recur in epilepsy is unknown. Recent genetic, pharmacological and electrophysiological data indicate a significant but undisclosed role for voltage-dependent calcium channels. Since the contribution such channels make to nerve function reflects the targeting of discrete subtypes to distinct cellular regions, we hypothesized that epilepsy reflects alterations in their spatiotemporal patterns of expression at the cell surface. To test this possibility, we examined the expression and distribution of hippocampal N-type calcium channels in an animal seizure model: kindling. Confocal microscopy of N-type calcium channels labeled with a new fluorescent ligand, coupled with a novel technique for analysing multiple images, revealed a 20-40% increase in their expression in CA1 and CA3 within 24 h post-seizure. These increases persisted in the dendritic fields of CA1, but had dissipated in CA3 by 28 days post-seizure. Such changes correlate poorly with cell number or synaptogenesis, but are consistent with increased N-type calcium channel expression on presynaptic terminals or, more likely, dendrites. These data rationalize recent electrophysiology and in situ hybridization data, and suggest that kindling alters N-type calcium channel trafficking mechanisms to cause a persistent, local, remodeling of their distributions in CA1 dendrites. The persistent induction of N-type calcium channels may be part of a mechanism for, and a hallmark of, synaptic plasticity, in which kindling represents a reinforcement of synapses en masse.

Animals↗

Substance P and neurokinin A mediate sensory synaptic transmission in young rat dorsal horn neurons.

Spinal nociceptive transmission is mediated by glutamate and neuropeptides such as substance P (SP) and neurokinin A (NKA). The neuropeptide-mediated excitatory postsynaptic potentials (EPSPs) had a slow onset and long duration. Here, we demonstrate SP- and NKA-mediated excitatory postsynaptic currents (EPSCs) in dorsal horn neurons of young rats using whole-cell patch-clamp recording techniques. After complete blockade of glutamate receptor-mediated currents, we observed a small residual EPSC. The residual EPSCs exhibited temporal summation in response to a train of stimulation (six pulses delivered at 10-50 Hz). High intensity stimulation (the same or greater than the stimulation threshold for nociceptive fibers in vivo) was required for evoking these summated EPSCs. Summated EPSCs were attenuated or abolished by capsaicin pretreatment, which depletes SP and NKA from presynaptic terminals; SP and NKA pretreatment; NK(1) or NK(2) receptor antagonists; and inhibition of postsynaptic G proteins. EPSCs were neither blocked by a metabotropic glutamate receptor antagonist nor a gamma-aminobutyric acid(B) receptor antagonist. The summated EPSCs were also sensitive to voltage-gated calcium channel antagonists or mu-opioid receptor activation by DAMGO. The present study provides electrophysiological evidence that suggests the possible contribution of SP and NKA to sensory synaptic transmission between primary afferent fibers and dorsal horn neurons.

2-Amino-5-phosphonovalerate↗

The effect of volatile anaesthetics on synaptic release and uptake of glutamate.

1. Volatile anaesthetics seem to exert their effects on several parts of the neuronal conducting system. 2. The effect on synaptic excitation seems to be quantitatively the most important (Berg-Johnsen and Langmoen, Acta Physiol. Scand. 128, 1986, 613-618) as 1 minimum alveolar concentration (MAC) of isoflurane reduces the activity in thin unmyelinated afferent fibres by 18%, excitatory synapses by 27% and postsynaptic neurones by 24%. 3. The reduction in excitatory synaptic transmission is caused by a decreased amount of transmitter glutamate in the synaptic cleft caused by a reduced release and increased uptake of glutamate in the presynaptic terminals (Larsen et al., Brain Res. 663, 1994, 335-337; Larsen et al., Br. J. Anaesth. 78, 1997, 55-59).

Anesthesia, Inhalation↗

Neuromuscular blocking activity of methanolic extract of Piper sarmentosum leaves in the rat phrenic nerve-hemidiaphragm preparation.

Methanolic extract of Piper sarmentosum Roxb. (Piperaceae) leaves was studied for the neuromuscular blocking activity in rat phrenic nerve-hemidiaphragm preparations. The plant extract, at concentrations of 3.2, 4.0, 4.8 and 6.4 mg/ml, exhibited an initially transient increase in twitch tension which was followed by a marked dose-related neurally-evoked twitch depression. The neuromuscular blocking effect produced by the plant extract was compared with d-tubocurarine (dTC) and succinylcholine (SCh). The EC50 for neurally-evoked twitch depression of the extract, dTC and SCh was 4.07 mg/ml, 1.1 microM and 15 microM, respectively. The neurally-evoked twitch depression produced by the extract was partially antagonized by tetraethylammonium (TEA) but not by neostigmine (NS). These findings suggested that the plant extract possessed a marked neuromuscular blocking activity at the neuromuscular junction and a possible mechanism which was likely to inhibit neurotransmitter (acetylcholine) release at the presynaptic terminal.

Action Potentials↗

Neuronal death in the central nervous system during development.

About half the neurons in the brain die at the time when their connections are being formed. This neuronal death is regulated by anterograde and retrograde signals that reflect both electrical activity and the uptake of trophic factors. Our recent data on the isthmo-optic projection indicate that there are in fact two different retrograde signals: a slow-acting survival signal mediated by a neurotrophin, and a fast-acting death signal mediated by calcium entry due to electrical activity in the presynaptic terminals. The developmental roles of the cell death are not well understood, but they appear to include the elimination of aberrant connections. The intracellular mechanisms of the cell death may not always correspond to the apoptotic ones so thoroughly investigated in vitro, because only one of the three morphological types occurring regularly in vivo resembles apoptosis. However, our experiments on retinal ganglion cells indicate that several apoptotic mechanisms apply in this particular in vivo situation: these include an involvement of oxygenated free radicals and glutathione, cell cycle-related events, and probably the synthesis of proteins promoting neuroprotection or cell death.

Animals↗

A model synapse that incorporates the properties of short- and long-term synaptic plasticity.

We propose a general computer model of a synapse, which incorporates mechanisms responsible for the realization of both short- and long-term synaptic plasticity-the two forms of experimentally observed plasticity that seem to be very significant for the performance of neuronal networks. The model consists of a presynaptic part based on the earlier 'double barrier synapse' model, and a postsynaptic compartment which is connected to the presynaptic terminal via a feedback, the sign and magnitude of which depend on postsynaptic Ca(2+) concentration. The feedback increases or decreases the amount of neurotransmitter which is in a ready for release state. The model adequately reproduced the phenomena of short- and long-term plasticity observed experimentally in hippocampal slices for CA3-CA1 synapses. The proposed model may be used in the investigation of certain real synapses to estimate their physiological parameters, and in the construction of realistic neuronal networks.

Animals↗

Sequence generation in arbitrary temporal patterns from theta-nested gamma oscillations: a model of the basal ganglia-thalamo-cortical loops.

A computational model that is able to generate sequences at arbitrary rates in a given serial order is presented for the cortico-basal ganglia (BG)-thalamic neural circuitry. Upon generating a sequence, this model stores information on the serial order of components in a cortical buffer by means of theta-nested gamma frequency oscillations observed experimentally in cortico-striatal neurons. This model assumes the existence of at least two functionally different classes of striatal spiny neurons. One class of striatal projection neurons (S-cells) select the first component in the cortical buffer through a temporal winner-take-all mechanism implemented by lateral inhibition. The inhibition should last for at least a few hundred milliseconds. In reality, it may be mediated by GABA(B) receptors at the presynaptic terminals of the cortico-striatal projection. The other class of striatal projection neurons (M-cells) retain the currently executed component in a cortico-BG-thalamic loop, for which the strong nonlinearity in transitions between up and down states of striatal neurons is crucial. For sequence generation at the level of striatum, the cortical neurons encoding the component selected for execution are inactivated by the feedback from the activated cortico-BG-thalamic loop. This model predicts that the transition to next component is triggered by a single external signal, i.e. the subthalamic input to the globus pallidum. This input gives a neural substrate for adjusting the rate of sequence generation.

Journal Article↗

Extrasynaptic glutamate spillover in the hippocampus: dependence on temperature and the role of active glutamate uptake.

At excitatory synapses on CA1 pyramidal cells of the hippocampus, a larger quantal content is sensed by N-methyl-D-aspartic acid receptors (NMDARs) than by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs). A novel explanation for this discrepancy is that glutamate released from terminals presynaptic to one cell can diffuse to and activate NMDARs, but not AMPARs, on a neighboring cell. If this occurs in the living brain, it could invalidate the view that glutamatergic synapses function as private communication channels between neurons. Here, we show that the discrepancy in quantal content mediated by the two receptors is greatly decreased at physiological temperature, compared with conventional recording conditions. This effect of temperature is not due to changes in release probability or uncovering of latent AMPARs. It is, however, partially reversed by the glutamate uptake inhibitor dihydrokainate. The results suggest that glutamate transporters play a critical role in limiting the extrasynaptic diffusion of glutamate, thereby minimizing cross-talk between neighboring excitatory synapses.

Animals↗

Calcium dependence of exocytosis and endocytosis at the cochlear inner hair cell afferent synapse.

Release of neurotransmitter at the inner hair cell (IHC) afferent synapse is a fundamental step in translating sound into auditory nerve excitation. To study the Ca2+ dependence of the underlying vesicle fusion and subsequent endocytosis, we combined Ca2+ uncaging with membrane capacitance measurements in mouse IHCs. Rapid elevations in [Ca2+]i above 8 microM caused a biphasic capacitance increase corresponding to the fusion of approximately 40,000 vesicles. The kinetics of exocytosis displayed a fifth-order Ca2+ dependence reaching maximal rates of >3 x 10(7) vesicle/s. Exocytosis was always followed by slow, compensatory endocytosis (tau congruent with 15 s). Higher [Ca2+]i increased the contribution of a faster mode of endocytosis with a Ca2+ independent time constant of approximately 300 ms. These properties provide for rapid and sustained transmitter release from this large presynaptic terminal.

Animals↗

NCAM and vesicle cycling: the importance of good glue in the long run.

Careful functional dissection of mouse neuromuscular junctions (NMJs) formed in the absence of NCAM reveal that in spite of relatively normal morphology and ultrastructure, the presynaptic terminals have profound alterations in the mechanisms of recycling of synaptic vesicles. These alterations, including the appearance of a brefeldin-sensitive pathway, leave synaptic transmission impaired for certain types of activity. NCAMs thus appear to play a critical role in the molecular organization and function of synaptic terminals.

Animals↗

Characterization of neocortical and hippocampal synaptosomes from temporal lobe epilepsy patients.

To investigate epilepsy-associated changes in the presynaptic terminal, we isolated and characterized synaptosomes from biopsies resected during surgical treatment of drug-resistant temporal lobe epilepsy (TLE) patients. Our main findings are: (1) The yield of synaptosomal protein from biopsies of epilepsy patients was about 25% of that from rat brain. Synaptosomal preparations were essentially free of glial contaminations. (2) Synaptosomes from TLE patients and naive rat brain, quickly responded to K(+)-depolarization with a 70% increase in intrasynaptosomal Ca(2+) ([Ca(2+)](i)), and a 40% increase in B-50/GAP-43 phosphorylation. (3) Neocortical and hippocampal synaptosomes from TLE patients contained 20-50% of the glutamate and gamma-aminobutyric acid (GABA) contents of rat cortical synaptosomes. (4) Although the absolute amount of glutamate and GABA released under basal conditions from neocortical synaptosomes of TLE patients was lower than from rat synaptosomes, basal release expressed as percentage of total content was higher (16.4% and 17.3%, respectively) than in rat (11.5% and 9. 9%, respectively). (5) Depolarization-induced glutamate and GABA release from neocortical synaptosomes from TLE patients was smaller than from rat synaptosomes (3.9% and 13.0% vs. 21.9% and 25.0%, respectively). (6) Analysis of breakdown of glial fibrillary acid protein (GFAP) indicates that resection time (anoxic period during the operation) is a critical parameter for the quality of the synaptosomes. We conclude that highly pure and viable synaptosomes can be isolated even from highly sclerotic human epileptic tissue. Our data show that in studies on human synaptosomes it is of critical importance to distinguish methodological (i.e., resection time) from pathology-related abnormalities.

Adult↗

Calcium oscillations encoding neuron-to-astrocyte communication.

The observation that the excitatory neurotransmitter glutamate released from presynaptic terminals can activate, beside the post-synaptic neuron, the glial cell astrocyte, stimulated glial cell research like no other event since the recognition in the 1980s that astrocytes can express on their membrane many receptors for classical neurotransmitters. The properties and the functional role(s) of such a neuron-to-astrocyte signaling have now become the focus of intense research in neurobiology. Indeed, a growing body of evidence has recently highlighted the ability of astrocytes to work as sophisticated detectors of synaptic activity: by changing the frequency of [Ca(2+)](i) oscillations evoked by the synaptic release of glutamate, these cells display the remarkable capacity to discriminate between different levels and patterns of synaptic activity. Furthermore, the observation that astrocytes increase the frequency of [Ca(2+)](i) oscillations in response to repetitive episodes of high neuronal activity challenges the common concept that memory function in the brain is an exclusive property of neuronal cells. Glutamate-mediated [Ca(2+)](i) elevations can also trigger in astrocytes the release of glutamate that can ultimately affect neuronal transmission. Given the wide role played by glutamate in brain physiology, our view on how the brain operates needs now to be revised taking into account the bi-directional, glutamatergic communication between neurons and astrocytes.

Animals↗

Histological and ultrastructural changes in the rat brain following systemic administration of picolinic acid.

Picolinic acid was administered intraperitoneally in a dose of 30, 60, or 100 mmol, once every 24 h for 8 days. Histologically, under normal conditions as well as when picolinic acid was administered in a dose of 30 mmol the brain formations exhibited characteristic features. When picolinic acid was administered in a dose of 60 mmol or 100 mmol, the alterations were profound and developed selectively in hippocampus, being much less intense in the substantia nigra and striatum. In such cases, injuries of neuronal cell bodies were accompanied by symptoms of spongiosis. Within the hippocampus, the neuronal cell body injury was selectively restricted to the hilar and CA3 regions of stratum pyramidale. Tissue spongiosis was more intense at the granular layer, particularly within the hilus and in the mossy fiber area at CA3. Histochemically, a variable intensity of the reaction of succinic and alpha-glycerophosphate dehydrogenases was demonstrated. A decrease in their activities was observed in areas where the neuronal cell body injuries and spongiosis took place. No changes in the Ca-ATP-ase activity in brain formation after picolinic acid treatment were observed. Ultrastructurally, the changes within substantia nigra were manifested by neuronal cell bodies of the dark type and dendritic degenerations. Also less damaged neuronal cell bodies were seen. They were swollen, depleted of polyribosomes with dilated elements of RER and altered mitochondria. Some of the dendritic profiles were swollen with lucent cytoplasm. Most of the boutons in synaptic contact zones were unchanged. Most presynaptic terminals which were in junction with dark dendrites were swollen with or without crystal-like aggregates of synaptic vesicles.

Animals↗

Imaging of catecholamine-secreting tumours: uses of MIBG in diagnosis and treatment.

MIBG radiolabelled with 131I or 123I is a radiopharmaceutical which is concentrated in neuroendocrine tumours, particularly phaeochromocytomas and neuroblastomas. This permits non-invasive whole-body scintigraphic screening for benign and malignant, familial and sporadic, intra-adrenal and extra-adrenal phaeochromocytomas and primary and metastatic neuroblastomas, with high sensitivity (85-90%) and specificity (> 95%). MIBG is also concentrated in presynaptic terminals of adrenergic, autonomically innervated organs such as the heart, and may be used as a non-invasive in vivo probe to study this system. Large doses of 131I-MIBG and 125I-MIBG have been used experimentally to selectively deliver therapeutic doses of radiotherapy to malignant phaeochromocytomas and refractory advanced neuroblastomas.

3-Iodobenzylguanidine↗