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Astrocytes are recognized as more important cells than historically thought in synaptic function through the reciprocal exchange of signaling with the neuronal synaptic elements. The idea that astrocytes are active elements in synaptic physiology is conceptualized in the Tripartite Synapse concept. This review article presents and discusses recent representative examples that highlight the heterogeneity of signaling in tripartite synapse function and its consequences on neural network function and animal behavior.

The synapse is densely packed with proteins involved in various highly regulated processes. Synaptic protein copy numbers and their stoichiometric distribution have a drastic influence on neuronal integrity and function. Therefore, the molecular analysis of synapses is a key element to understand their architecture and function. The overall structure of the synapse has been revealed with an exquisite amount of details by electron microscopy. However, the molecular composition and the localization of proteins are more easily addressed with fluorescence imaging, especially with the improved resolution achieved by super-resolution microscopy techniques. Notably, the fast improvement of imaging instruments has not been reflected in the optimization of biological sample preparation. During recent years, large efforts have been made to generate affinity probes smaller than conventional antibodies adapted for fluorescent super-resolution imaging. In this review, we briefly discuss the current views on synaptic organization and necessary key technologies to progress in the understanding of synaptic physiology. We also highlight the challenges faced by current fluorescent super-resolution methods, and we describe the prerequisites for an ideal study of synaptic organization.

Cortical information processing at the cellular level has predominantly been studied in local networks, which are dominated by strong vertical connectivity between layers. However, recent studies suggest that the bulk of axons targeting pyramidal neurons most likely originate from outside this local range, emphasizing the importance of horizontal connections. We mapped a subset of these connections to L5B pyramidal neurons in rat somatosensory cortex with photostimulation, identifying intact projections up to a lateral distance of 2 mm. Our estimates of the spatial distribution of cells presynaptic to L5B pyramids support the idea that the majority is located outside the local volume. The synaptic physiology of horizontal connections does not differ markedly from that of local connections, whereas the layer and cell-type-dependent pattern of innervation does. Apart from L2/3, L6A provides a strong source of horizontal connections. Implementing our data into a spiking neuronal network model shows that more horizontal connections promote robust asynchronous ongoing activity states and reduce noise correlations in stimulus-induced activity. QC 20141204

Studying synaptic physiology remains an important endeavor for understanding the mechanisms that underlie neurotransmitter release as well as synaptic development and refinement. It is also critical to understanding brain function. The basic conceptual framework for synaptic physiology was worked out by Katz and colleagues and has coalesced around three factors: the number of releasable vesicles (N), the probability of release (P), and quantal size (Q). Here, I discuss a few key experiments related to these quantities and how they are assessed.

This protocol describes how to prepare brain slices for electrophysiology, with an emphasis on synaptic physiology in voltage clamp. This approach remains the gold standard for understanding the properties of individual neurons, as well as the connections between neurons.

Synapses exhibit diverse functional properties, and it seems likely that these properties are specialized to perform specific computations. The Drosophila antennal lobe provides a useful experimental preparation for exploring the relationship between synaptic physiology and neural computations. This review summarizes recent progress in describing synaptic properties in the Drosophila antennal lobe. These studies reveal that several types of synapses in this circuit are highly specialized, and that these specializations are in some cases under tight regulatory control. These synaptic specializations can be understood in terms of the computational features they confer on the circuit. Specifically, many of these properties appear to promote odor detection when odor concentrations are low, while promoting adaptive gain control when odor concentrations are high.

Glial cells are instrumental for many aspects of nervous-system function. Interestingly, complex neuron—glial interactions at synapses are commonly found in both invertebrates and vertebrates. Although these interactions are known to be important for synaptic physiology, the cellular processes and molecular mechanisms involved have not been fully uncovered. Identifying the common and unique features of neuron—glial interactions between invertebrates and vertebrates may provide valuable insights into the relationship of neuron—glial cross-talk to nervous-system function. This review highlights selected studies that have revealed structural and functional insights into neuron—glial interactions at synapses in invertebrate and vertebrate model systems. NEUROSCIENTIST 13(6): 657—666, 2007. DOI: 10.1177/1073858407304393

Nucleus angularis (NA), one of the two cochlear nuclei in birds, is important for processing sound intensity for localization and most likely has role in sound recognition and other auditory tasks. Because the synaptic properties of auditory nerve inputs to the cochlear nuclei are fundamental to the transformation of auditory information, we studied the properties of these synapses onto NA neurons using whole cell patch-clamp recordings from auditory brain stem slices from embryonic chickens (E16-E20). We measured spontaneous excitatory postsynaptic currents (EPSCs), and evoked EPSCs and excitatory postsynaptic potentials (EPSPs) by using extracellular stimulation of the auditory nerve. These excitatory EPSCs were mediated by AMPA and N-methyl-D-aspartate (NMDA) receptors. The spontaneous EPSCs mediated by AMPA receptors had submillisecond decay kinetics (556 micros at E19), comparable with those of other auditory brain stem areas. The spontaneous EPSCs increased in amplitude and became faster with developmental age. Evoked EPSC and EPSP amplitudes were graded with stimulus intensity. The average amplitude of the EPSC evoked by minimal stimulation was twice as large as the average spontaneous EPSC amplitude (approximately 110 vs. approximately 55 pA), suggesting that single fibers make multiple contacts onto each postsynaptic NA neuron. Because of their small size, minimal EPSPs were subthreshold, and we estimate at least three to five inputs were required to reach threshold. In contrast to the fast EPSCs, EPSPs in NA had a decay time constant of approximately 12.5 ms, which was heavily influenced by the membrane time constant. Thus NA neurons spatially and temporally integrate auditory information arriving from multiple auditory nerve afferents.

Kainate receptors (KARs) play an important role in synaptic physiology, plasticity, and pathological phenomena such as epilepsy. However, the physiological implications for neuronal networks of the distinct expression patterns of KAR subunits are unknown. Using KAR knock-out mice, we show that subunits glutamate receptor (GluR) 5 and GluR6 play distinct roles in kainate-induced gamma oscillations and epileptiform burst activity. Ablation of GluR5 leads to a higher susceptibility of the network to the oscillogenic and epileptogenic effects of kainate, whereas lack of GluR6 prevents kainate-induced gamma oscillations or epileptiform bursts. Based on experimental and simulated neuronal network data as well as the consequences of GluR5 and GluR6 expression for cellular and synaptic physiology, we propose that the functional interplay of GluR5-containing KARs on axons of interneurons and GluR6-containing KARs in the somatodendritic region of both interneurons and pyramidal cells underlie the oscillogenic and epileptogenic effects of kainate.

Synapses are the fundamental units of information processing in the mammalian brain. Much of our understanding of their functional properties comes from voltage-clamp analysis, the predominant approach for investigating synaptic physiology. Here, we reveal that voltage clamp is completely ineffective for most excitatory synapses due to spine electrical compartmentalization. Under local dendritic voltage clamp, single-spine activation produced large spine head depolarizations that severely distorted measurements and recruited voltage-dependent channels. To overcome these voltage-clamp errors, we developed an approach to provide new, accurate measurements of synaptic conductance. Single-synapse AMPA conductance was much larger than previously appreciated, producing saturation effects on synaptic currents. We conclude that electrical compartmentalization profoundly shapes both synaptic function and how that function can be assessed with electrophysiological methods.

Each stage of the striate cortical circuit extracts novel information about the visual environment. We asked if this analytic process reflected laminar variations in synaptic physiology by making whole-cell recording with dye-filled electrodes from the cat's visual cortex and thalamus; the stimuli were flashed spots. Thalamic afferents terminate in layer 4, which contains two types of cell, simple and complex, distinguished by the spatial structure of the receptive field. Previously, we had found that the postsynaptic and spike responses of simple cells reliably followed the time course of flash-evoked thalamic activity. Here we report that complex cells in layer 4 (or cells intermediate between simple and complex) similarly reprised thalamic activity (response/trial, 99 +/- 1.9 %; response duration 159 +/- 57 ms; latency 25 +/- 4 ms; average +/- standard deviation; n = 7). Thus, all cells in layer 4 share a common synaptic physiology that allows secure integration of thalamic input. By contrast, at the second cortical stage (layer 2+3), where layer 4 directs its output, postsynaptic responses did not track simple patterns of antecedent activity. Typical responses to the static stimulus were intermittent and brief (response/trial, 31 +/- 40 %; response duration 72 +/- 60 ms, latency 39 +/- 7 ms; n = 11). Only richer stimuli like those including motion evoked reliable responses. All told, the second level of cortical processing differs markedly from the first. At that later stage, ascending information seems strongly gated by connections between cortical neurons. Inputs must be combined in newly specified patterns to influence intracortical stages of processing.

The concept of the tripartite synapse reflects the important role that astrocytic processes are thought to play in the function and regulation of neuronal synapses in the mammalian nervous system. However, many basic aspects regarding the dynamic interplay between pre- and postsynaptic neuronal structures and their astrocytic partners remain to be explored. A major experimental hurdle has been the small physical size of the relevant glial and synaptic structures, leaving them largely out of reach for conventional light microscopic approaches such as confocal and two-photon microscopy. Hence, most of what we know about the organization of the tripartite synapse is based on electron microscopy, which does not lend itself to investigating dynamic events and which cannot be carried out in parallel with functional assays. The development and application of superresolution microscopy for neuron–glia research is opening up exciting experimental opportunities in this regard. In this paper, we provide a basic explanation of the theory and operation of stimulated emission depletion (STED) microscopy, outlining the potential of this recent superresolution imaging modality for advancing our understanding of the morpho-functional interactions between astrocytes and neurons that regulate synaptic physiology.

A key question in synaptic physiology is what determines the release probability of a synaptic vesicle. In this issue of Neuron , Wadel et al. shed UV light on this problem, finding that hard-to-release vesicles are too far from Ca 2+ channels.

Baraban, Scott and Philip A. Schwartzkroin. Effects of prenatal cocaine exposure on the developing hippocampus: intrinsic and synaptic physiology. J. Neurophysiol. 77: 126–136, 1977. A variety of neurological complications has been reported in infants exposed to cocaine during gestation. In the present study, intrinsic cell properties of hippocampal neurons from CA1, CA3, and dentate gyrus regions were measured and compared in tissue from neonatal rats exposed to saline or cocaine in utero. Synaptic properties of the CA1 pyramidal cell region were analyzed at postnatal day (P) 20 with the use of extracellular and intracellular recording techniques. In vitro intracellular recordings ( n = 223) obtained at P10, P15 and P20 in tissue from cocaine- and saline-exposed animals revealed no differences in standard cell properties such as resting membrane potential, input resistance, time constant, and action potential amplitude or duration. Hippocampal slices from cocaine-exposed animals exhibited a marked reduction of spike frequency adaptation for all three types of principal hippocampal neurons (e.g., CA1, CA3, and granule cells). The amplitudes of afterhyperpolarizations following a spike train were also decreased in CA1 and CA3 cells in tissue from cocaine-exposed animals. Extracellular and intracellular recordings in the CA1 pyramidal cell region at P20 were obtained to assess and compare synaptic function in tissue from cocaine- and saline-exposed animals. In hippocampal slices from cocaine-exposed animals, synaptic responses in the CA1 region were characterized by multiple population spike activity and reduced inhibitory postsynaptic potentials. The reduction in fast inhibitory postsynaptic potential conductance was not associated with a change in reversal potential. These results suggest that gestational cocaine exposure induces significant changes in intrinsic and synaptic electrophysiological properties of hippocampal neurons in the developing animal. The cell and synaptic features are consistent with an increase in hippocampal excitability, which may contribute to the neurobehavioral deficits and epileptogenic predisposition reported in this infant population. As such, this in utero drug exposure model may provide a useful system in which to elucidate and study the basic cellular mechanisms underlying neurological complications associated with maternal cocaine abuse.

Fly and vertebrate nervous systems share many organizational features, such as layers, columns and glomeruli, and utilize similar synaptic components, such as ion channels and receptors. Both also exhibit similar network features. Recent technological advances, especially in electron microscopy, now allow us to determine synaptic circuits and identify pathways cell-by-cell, as part of the fly's connectome. Genetic tools provide the means to identify synaptic components, as well as to record and manipulate neuronal activity, adding function to the connectome. This review discusses technical advances in these emerging areas of functional connectomics, offering prognoses in each and identifying the challenges in bridging structural connectomics to molecular biology and synaptic physiology, thereby determining fundamental mechanisms of neural computation that underlie behavior.

Neurons in the sensory system exhibit changes in excitability that unfold over many time scales. These fluctuations produce noise and could potentially lead to perceptual errors. However, to prevent such errors, postsynaptic neurons and synapses can adapt and counteract changes in the excitability of presynaptic neurons. Here we ask how neurons could optimally adapt to minimize the influence of changing presynaptic neural properties on their outputs. The resulting model, based on Bayesian inference, explains a range of physiological results from experiments which have measured the overall properties and detailed time-course of sensory tuning curve adaptation in the early visual cortex. We show how several experimentally measured short term plasticity phenomena can be understood as near-optimal solutions to this adaptation problem. This framework provides a link between high level computational problems, the properties of cortical neurons, and synaptic physiology., National Eye Institute (EY01362), Chicago Community Trust, National Institutes of Health (U.S.) (1R01NS063399 ), National Institutes of Health (U.S.) (2P01NS044393)

Neurons in plastic regions of the brain undergo fundamental changes in the number of cells connecting to them as a result of development, plasticity and disease. Across these same time periods, functional changes in cellular and synaptic physiology are known to occur and are often characterized as developmental features of these periods. However, it remains possible that many such changes are direct consequences of the modified degree of partnering, and that neurons intrinsically scale their physiological parameters with network size. To systematically vary a recurrent network's number of neurons while measuring its synaptic properties, we used microfabricated extracellular matrix adhesive islands created with soft lithography to culture neuronal clusters of precise sizes, and assessed their intrinsic connectivity using intracellular recordings and confocal microscopy. Both large and small clusters supported constant densities of excitatory and inhibitory neurons. However, neurons that were provided with more potential partners (larger clusters) formed more connections per cell via an expanded dendritic surface than cocultured smaller clusters. Electrophysiologically, firing rate was preserved across clusters even as size and synapse number increased, due in part to synapses in larger networks having reduced unitary strengths, and sparser paired connectivity. Larger networks also featured a particular increase in the number of excitatory connections onto inhibitory dendrites. We suggest that these specific homeostatic mechanisms, which match the number, strength, and architecture of connections to the number of total available cellular partners in the network, could account for several known phenomena implicated in the formation, organization and degeneration of neuronal circuits.

When given orally to a transgenic mouse model of Alzheimer disease, cyclohexanehexol stereoisomers inhibit aggregation of amyloid beta peptide (Abeta) into high-molecular-weight oligomers in the brain and ameliorate several Alzheimer disease-like phenotypes in these mice, including impaired cognition, altered synaptic physiology, cerebral Abeta pathology and accelerated mortality. These therapeutic effects, which occur regardless of whether the compounds are given before or well after the onset of the Alzheimer disease-like phenotype, support the idea that the accumulation of Abeta oligomers has a central role in the pathogenesis of Alzheimer disease.

Recent studies suggest that endogenous neurotrophins play a central role in the patterning of cortical connections and in cortical synaptic physiology. Do these effects of neurotrophins reflect independent cellular events, or are they manifestations of a single cellular mechanism central to developmental plasticity?

Nguyen, Peter V., Leo Marin, and Harold L. Atwood. Synaptic physiology and mitochondrial function in crayfish tonic and phasic motor neurons. J. Neurophysiol. 78: 281–294, 1997. Phasic and tonic motor neurons of crustaceans differ strikingly in their junctional synaptic physiology. Tonic neurons generally produce small excitatory postsynaptic potentials (EPSPs) that facilitate strongly as stimulation frequency is increased, and normally show no synaptic depression. In contrast, phasic neurons produce relatively large EPSPs with weak frequency facilitation and pronounced depression. We addressed the hypothesis that mitochondrial function is an important determinant of the features of synaptic transmission in these neurons. Mitochondrial fluorescence was measured with confocal microscopy in phasic and tonic axons and terminals of abdominal and leg muscles after exposure to supravital mitochondrial fluorochromes, rhodamine-123 (Rh123) and 4-diethylaminostyryl-N-methylpyridinium iodide (4-Di-2-Asp). Mitochondria of tonic axons and neuromuscular junctions had significantly higher mean Rh123 and 4-Di-2-Asp fluorescence than in phasic neurons, indicating more accumulation of the fluorochromes. Mitochondrial membrane potential, which is responsible for Rh123 uptake and is related to mitochondrial oxidative activity (the production of ATP by oxidation of metabolic substrates), is likely higher in tonic axons. Electron microscopy showed that tonic axons contain approximately fivefold more mitochondria per μm2cross-sectional area than phasic axons. Neuromuscular junctions of tonic axons also have a much higher mitochondrial content than those of phasic axons. We tested the hypothesis that synaptic fatigue resistance is dependent on mitochondrial function in crayfish motor axons. Impairment of mitochondrial function by uncouplers of oxidative phosphorylation, dinitrophenol or carbonyl cyanide m-chlorophenylhydrazone, or by the electron transport inhibitor sodium azide, led to marked synaptic depression of a tonic axon and accelerated depression of a phasic axon during maintained stimulation. Iodoacetate, an inhibitor of glycolysis, and chloramphenicol, a mitochondrial protein synthesis inhibitor, had no significant effects on either mitochondrial fluorescence or synaptic depression in tonic or phasic axons. Collectively, the results provide evidence that mitochondrial oxidative metabolism is important for sustaining synaptic transmission during maintained stimulation of tonic and phasic motor neurons. Tonic neurons have a higher mitochondrial content and greater oxidative activity; these features are correlated with their greater resistance to synaptic depression. Conversely, phasic neurons have a lower mitochondrial content, less oxidative activity, and greater synaptic fatigability.

Do graded potential synapses work the same way as action potential synapses? Recent work emphasizes the differences and suggests that graded potential synapses are not all the same either.

Slices of the field CA1 minus the stratum moleculare were prepared from the rat hippocampus and maintained in an in vitro recording chamber. The physiological properties of these "minislices" were similar to those reported for non-dissected hippocampal slices. Slices receiving various patterns of electrical stimulation through multiple electrodes were subsequently homogenized and crude membrane fractions prepared; the binding of [3H]glutamate was measured by a rapid filtration assay. Binding in membranes prepared from control slices exhibited kinetic properties and sensitivities to pharmacological and ionic manipulations which were comparable to those found in previous studies using conventional fractionation and assay techniques. Brief bursts of high-frequency stimulation increased [3H]glutamate binding compared to non-stimulated controls in 3 separate experiments. Stimulation at low frequency or at high frequency in low calcium medium did not produce this effect. In addition to suggesting that glutamate binding sites are regulated by patterns of afferent activity, these findings indicate that the minislice preparation should be of general utility in relating synaptic physiology to synaptic chemistry.

The complexity and relative inaccessibility of the central nervous system have greatly impeded the study of synaptic physiology. However, in recent years a fruitful approach to the problem has been to apply methods of chemi­ cal and pharmacological analysis. The present review is almost exclusively limited to biochemical studies of synaptic function in the mammalian eNS. It is not a comprehensive review of this vast topic, but rather an attempt to highlight important current approaches and trends. Previous reviewers have dealt with many aspects of synaptic biochemistry and pharmacology (9, 64, 66, 68,73, 80,92,95, 103, 121 , 142,1 58, 163,173). The topics reviewed in the present paper include general metabolism of synapses and the more spe­ cialized presynaptic metabolism of neurotransmitters as well as their post­ synaptic actions.