Localization and mobility of omega-conotoxin-sensitive Ca2+ channels in hippocampal CA1 neurons

Voltage-dependent Ca2+ channels (VDCCs) are modulators of synaptic plasticity, oscillatory behavior, and rhythmic firing in brain regions such as the hippocampus. The distribution and lateral mobility of VDCCs on CA1 hippocampal neurons have been determined with biologically active fluorescent and biotinylated derivatives of the selective probe omega-conotoxin in conjunction with circular dityndallism, digital fluorescence imaging, and photobleach recovery microscopy. On noninnervated cell bodies, VDCCs were found to be organized in multiple clusters, whereas after innervation the VDCCs were concentrated and immobilized at synaptic contact sites. On dendrites, VDCC distribution was punctate and was interrupted by extensive bare regions or abruptly terminated. More than 85% of the dendritic VDCCs were found to be immobile by fluorescence photobleach recovery. Thus, before synaptic contact, specific mechanisms target, segregate, and immobilize VDCCs to neuronal cell bodies and to specialized dendritic sites. Regulation of this distribution may be critical in determining the firing activity and integrative properties of hippocampal CA1 neurons.     

Rapid increase in clusters of presynaptic proteins at onset of long-lasting potentiation

A change in the efficiency of synaptic communication between neurons is thought to underlie learning. Consistent with recent studies of such changes, we find that long-lasting potentiation of synaptic transmission between cultured hippocampal neurons is accompanied by an increase in the number of clusters of postsynaptic glutamate receptors containing the subunit GluR1. In addition, potentiation is accompanied by a rapid and long-lasting increase in the number of clusters of the presynaptic protein synaptophysin and the number of sites at which synaptophysin and GluR1 are colocalized. These results suggest that potentiation involves rapid coordinate changes in the distribution of proteins in the presynaptic neuron as well as the postsynaptic neuron.     

Uptake of glutamate into synaptic vesicles by an inorganic phosphate transporter

Previous work has identified two families of proteins that transport classical neurotransmitters into synaptic vesicles, but the protein responsible for vesicular transport of the principal excitatory transmitter glutamate has remained unknown. We demonstrate that a protein that is unrelated to any known neurotransmitter transporters and that was previously suggested to mediate the Na(+)-dependent uptake of inorganic phosphate across the plasma membrane transports glutamate into synaptic vesicles. In addition, we show that this vesicular glutamate transporter, VGLUT1, exhibits a conductance for chloride that is blocked by glutamate.     

The connectome of a decision-making neural network

In order to understand the nervous system, it is necessary to know the synaptic connections between the neurons, yet to date, only the wiring diagram of the adult hermaphrodite of the nematode Caenorhabditis elegans has been determined. Here, we present the wiring diagram of the posterior nervous system of the C. elegans adult male, reconstructed from serial electron micrograph sections. This region of the male nervous system contains the sexually dimorphic circuits for mating. The synaptic connections, both chemical and gap junctional, form a neural network with four striking features: multiple, parallel, short synaptic pathways directly connecting sensory neurons to end organs; recurrent and reciprocal connectivity among sensory neurons; modular substructure; and interneurons acting in feedforward loops. These features help to explain how the network robustly and rapidly selects and executes the steps of a behavioral program on the basis of the inputs from multiple sensory neurons.     

Sustained dendritic gradients of Ca2+ induced by excitatory amino acids in CA1 hippocampal neurons

Spatially resolved measurements of intracellular free calcium and of the changes produced by excitatory amino acids were made in neurons isolated from adult mammalian brain. Extremely long-lasting (minutes) Ca2+ gradients were induced in the apical dendrites of hippocampal CA1 neurons after brief (1 to 3 seconds), local application of either glutamate or N-methyl-D-aspartate (NMDA). These gradients reflect the continuous flux of Ca2+ into the dendrite. The sustained gradients, but not the immediate transient response to the agonists, were prevented by prior treatment with the protein kinase C inhibitor sphingosine. Expression of the long-lasting Ca2+ gradients generally required a priming or conditioning stimulus with the excitatory agonist. The findings demonstrate a coupling between NMDA receptor activation and long-lasting intracellular Ca2+ elevation that could contribute to certain use-dependent modifications of synaptic responses in hippocampal CA1 neurons.     

Asymmetrical allocation of NMDA receptor epsilon2 subunits in hippocampal circuitry

Despite its implications for higher order functions of the brain, little is currently known about the molecular basis of left-right asymmetry of the brain. Here we report that synaptic distribution of N-methyl-D-aspartate (NMDA) receptor GluRepsilon2 (NR2B) subunits in the adult mouse hippocampus is asymmetrical between the left and right and between the apical and basal dendrites of single neurons. These asymmetrical allocations of epsilon2 subunits differentiate the properties of NMDA receptors and synaptic plasticity between the left and right hippocampus. These results provide a molecular basis for the structural and functional asymmetry of the mature brain.     

New insights into neuron-glia communication

Two-way communication between neurons and nonneural cells called glia is essential for axonal conduction, synaptic transmission, and information processing and thus is required for normal functioning of the nervous system during development and throughout adult life. The signals between neurons and glia include ion fluxes, neurotransmitters, cell adhesion molecules, and specialized signaling molecules released from synaptic and nonsynaptic regions of the neuron. In contrast to the serial flow of information along chains of neurons, glia communicate with other glial cells through intracellular waves of calcium and via intercellular diffusion of chemical messengers. By releasing neurotransmitters and other extracellular signaling molecules, glia can affect neuronal excitability and synaptic transmission and perhaps coordinate activity across networks of neurons.     

Clusters of hyperactive neurons near amyloid plaques in a mouse model of Alzheimer's disease

The neurodegeneration observed in Alzheimer's disease has been associated with synaptic dismantling and progressive decrease in neuronal activity. We tested this hypothesis in vivo by using two-photon Ca2+ imaging in a mouse model of Alzheimer's disease. Although a decrease in neuronal activity was seen in 29% of layer 2/3 cortical neurons, 21% of neurons displayed an unexpected increase in the frequency of spontaneous Ca2+ transients. These "hyperactive" neurons were found exclusively near the plaques of amyloid beta-depositing mice. The hyperactivity appeared to be due to a relative decrease in synaptic inhibition. Thus, we suggest that a redistribution of synaptic drive between silent and hyperactive neurons, rather than an overall decrease in synaptic activity, provides a mechanism for the disturbed cortical function in Alzheimer's disease.     

Neurobiology. Long-sought protein packages glutamate

Glutamate, one of two neurotransmitters that send the basic "stop" and "go" signals that most other neurotransmitters merely modulate, is called into action wherever rapid-fire excitatory signals are needed--say, for vision or learning. For decades, researchers have been looking for the protein that packages glutamate for express delivery to other neurons. On page 957, researchers report that they've found this elusive transporter, dubbed VGLUT1.     

Synaptic transmission between dissociated adult mammalian neurons and attached synaptic boutons

In most studies of synaptic currents in mammalian central neurons, preparations have been used in which synaptic currents are recorded at some distance from the synapse itself. This procedure introduces problems in interpretation of the kinetics and voltage-dependent properties of the synaptic current. These problems have now been overcome by the development of a preparation in which presynaptic vesicle-containing boutons have been coisolated with the soma of individual neurons, thus providing the opportunity to study synaptic currents under conditions of both adequate voltage control and internal ionic perfusion. Spontaneous synaptic currents mediated by gamma-aminobutyric acid and excitatory amino acids were recorded from neurons isolated from a mammalian medial solitary tract nucleus. Calcium- and depolarization-dependent spontaneous currents of several to hundreds of picoamperes occurred with rapid rise times of 0.8 to 3 milliseconds and decays at least ten times as long.     

Induction of a neuronal proteoglycan by the NMDA receptor in the developing spinal cord

Activation of the N-methyl-D-aspartate (NMDA) subclass of glutamate receptors is a critical step in the selection of appropriate synaptic connections in the developing visual systems of cat and frog. Activity-dependent development of mammalian motor neurons was shown to be similarly mediated by activation of the NMDA receptor. The expression of the Cat-301 proteoglycan on motor neurons was developmentally regulated and could be specifically inhibited by blockade of the NMDA receptor at the spinal segmental level. In the adult, Cat-301 immunoreactivity on motor neurons was not diminished by NMDA receptor blockade. The NMDA receptor may regulate the expression of a class of neuronal proteins (of which Cat-301 is one example) that underlie the morphological and physiological features of activity-dependent development.     

GABAergic神经元在BARREL和VPM区组织结构及形态特点的研究（英文）

To understand the structure of GABAergic neurons in the VMP and "barrel",the distribution of GABAergic neurons in the two areas were studied through immunohistochemistry and Laser Scanning Confocal Microscope.The results show that the distribution of GABAergic neurons in VMP and barrel are different,and the coding of information transmission in the two areas are also dissimilar;GABAergic neurons mainly distribute among the lines asymmetrically in VMP,the somata,dendrite and axon of GABAergic neurons are restricted in the "barrel",rarely having synaptic connections with other "barrel" around.Therefore,VMP and barrel may have different roles in transmission and on processing of informatiton.     

Temporal specificity in the cortical plasticity of visual space representation

The circuitry and function of mammalian visual cortex are shaped by patterns of visual stimuli, a plasticity likely mediated by synaptic modifications. In the adult cat, asynchronous visual stimuli in two adjacent retinal regions controlled the relative spike timing of two groups of cortical neurons with high precision. This asynchronous pairing induced rapid modifications of intracortical connections and shifts in receptive fields. These changes depended on the temporal order and interval between visual stimuli in a manner consistent with spike timing-dependent synaptic plasticity. Parallel to the cortical modifications found in the cat, such asynchronous visual stimuli also induced shifts in human spatial perception.     

GABAergic神经元在BARREL和VPM区组织结构及形态特点的免疫组织化学研究

[目的]研究GABAergic神经元在VPM和"barrel"区的组织结构及形态特点;[方法]通过免疫组织化学的方法和激光共聚焦电子显微镜研究GABAergic神经元在VPM和"barrel"区分布状态;[结果]GABAergic神经元在VPM和"barrel"区分布状态不同,信息传递这2个区域编码程度也不一样;GABAergic在VPM区主要分布在列与列之间,且呈非对称分布,而GABAergic神经元的胞体、树突和轴突出现限定在"barrel"内,与周围"barrel"很少形成突触联系。[结论]提示VPM和"barrel"可能在信息传递及处理过程中具有不同的功能。     

Spine-type-specific recruitment of newly synthesized AMPA receptors with learning

The stabilization of long-term memories requires de novo protein synthesis. How can proteins, synthesized in the soma, act on specific synapses that participate in a given memory? We studied the dynamics of newly synthesized AMPA-type glutamate receptors (AMPARs) induced with learning using transgenic mice expressing the GluR1 subunit fused to green fluorescent protein (GFP-GluR1) under control of the c-fos promoter. We found learning-associated recruitment of newly synthesized GFP-GluR1 selectively to mushroom-type spines in adult hippocampal CA1 neurons 24 hours after fear conditioning. Our results are consistent with a "synaptic tagging" model to allow activated synapses to subsequently capture newly synthesized receptor and also demonstrate a critical functional distinction in the mushroom spines with learning.     

Adenosine 3',5'-monophosphate: electrophysiological evidence for a role in synaptic transmission

Synaptic potentials and changes in resting membrane potentials of superior cervical ganglia of the rabbit were measured in the presence of adenosine 3',5'-monophosphate and agents that affect its metabolism. Adenosine 3',5'-monophosphate and its mono- and dibutyryl derivatives caused a hyperpolarization of the postganglionic neurons. Theophylline potentiated the slow inhibitory postsynaptic potential that follows synaptic transmission, as well as the hyperpolarization of postganglionic neurons caused by exogenous dopamine. Conversely, prostaglandin E(1) inhibited both the slow inhibitory postsynaptic potential and the dopamine-induced hyperpolarization. We hypothesize that the slow inhibitory postsynaptic potential as well as the dopamine-induced hyperpolarization result from increased amounts of adenosine 3'5'-monophosphate in the postganglionic neurons. The dibutyryl derivative of guanosine 3'5'-monophosphate caused a depolarization of the postganglionic neurons, which is consistent with the possibility that guanosine 3'5'-monophosphate mediates synaptic transmission at muscarinic cholinergic synapses.     

Androgens regulate the dendritic length of mammalian motoneurons in adulthood

Sex steroid hormones have been thought to alter behaviors in adulthood by changing the activity of neural circuits rather than by inducing major structural changes in these pathways. In a group of androgen-sensitive motoneurons that mediate male copulatory functions, decreases in androgen levels after castration of adult rats produced dramatic structural changes, decreasing both the dendritic length and soma size of these motoneurons. These changes were reversed by androgen replacement. These results imply a surprising degree of synaptic plasticity in adult motoneurons and suggest that normal changes in androgen levels in adulthood are associated with significant alterations in the structure and function of these neurons.     

Experience strengthening transmission by driving AMPA receptors into synapses

The mechanisms underlying experience-dependent plasticity in the brain may depend on the AMPA subclass of glutamate receptors (AMPA-Rs). We examined the trafficking of AMPA-Rs into synapses in the developing rat barrel cortex. In vivo gene delivery was combined with in vitro recordings to show that experience drives recombinant GluR1, an AMPA-R subunit, into synapses formed between layer 4 and layer 2/3 neurons. Moreover, expression of the GluR1 cytoplasmic tail, a construct that inhibits synaptic delivery of endogenous AMPA-Rs during long-term potentiation, blocked experience-driven synaptic potentiation. In general, synaptic incorporation of AMPA-Rs in vivo conforms to rules identified in vitro and contributes to plasticity driven by natural stimuli in the mammalian brain.     

Target cell-dependent normalization of transmitter release at neocortical synapses

The efficacy and short-term modification of neocortical synaptic connections vary with the type of target neuron. We investigated presynaptic Ca2+ and release probability at single synaptic contacts between pairs of neurons in layer 2/3 of the rat neocortex. The amplitude of Ca2+ signals in boutons of pyramids contacting bitufted or multipolar interneurons or other pyramids was dependent on the target cell type. Optical quantal analysis at single synaptic contacts suggested that release probabilities are also target cell-specific. Both the Ca2+ signal and the release probability of different boutons of a pyramid contacting the same target cell varied little. We propose that the mechanisms that regulate the functional properties of boutons of a pyramid normalize the presynaptic Ca2+ influx and release probability for all those boutons that innervate the same target cell.     

From stimulation to undulation: a neuronal pathway for the control of swimming in the leech

Initiation and performance of the swimming movement in the leech (Hirudo medicinalis) are controlled by neurons organized at at least four functional levels-sensory neurons, gating neurons, oscillator neurons, and motor neurons. A paired neuron, designated as Trl, in the subesophageal ganglion of the leech has now been shown to define a fifth level, interposed between sensory and gating neurons. Cell Trl is activated by pressure and nociceptive mechanosensory neurons, which mediate bodywall stimulus-evoked swimming activity in intact leeches. In the isolated leech nervous system, brief stimulation of cell Trl elicits sustained activation of the gating neurons and triggers the onset of swimmning activity. The synaptic interactions between all five levels of control are direct. Discovery of the Trl cells thus completes the identification of a synaptic pathway by which mechanosensory stimulation leads to the swimming movements of the leech.