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      Determining the Neurotransmitter Concentration Profile at Active Synapses

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          Abstract

          Establishing the temporal and concentration profiles of neurotransmitters during synaptic release is an essential step towards understanding the basic properties of inter-neuronal communication in the central nervous system. A variety of ingenious attempts has been made to gain insights into this process, but the general inaccessibility of central synapses, intrinsic limitations of the techniques used, and natural variety of different synaptic environments have hindered a comprehensive description of this fundamental phenomenon. Here, we describe a number of experimental and theoretical findings that has been instrumental for advancing our knowledge of various features of neurotransmitter release, as well as newly developed tools that could overcome some limits of traditional pharmacological approaches and bring new impetus to the description of the complex mechanisms of synaptic transmission.

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          Most cited references126

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          High-density mapping of single-molecule trajectories with photoactivated localization microscopy.

          We combined photoactivated localization microscopy (PALM) with live-cell single-particle tracking to create a new method termed sptPALM. We created spatially resolved maps of single-molecule motions by imaging the membrane proteins Gag and VSVG, and obtained several orders of magnitude more trajectories per cell than traditional single-particle tracking enables. By probing distinct subsets of molecules, sptPALM can provide insight into the origins of spatial and temporal heterogeneities in membranes.
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            Enforcement of temporal fidelity in pyramidal cells by somatic feed-forward inhibition.

            The temporal resolution of neuronal integration depends on the time window within which excitatory inputs summate to reach the threshold for spike generation. Here, we show that in rat hippocampal pyramidal cells this window is very narrow (less than 2 milliseconds). This narrowness results from the short delay with which disynaptic feed-forward inhibition follows monosynaptic excitation. Simultaneous somatic and dendritic recordings indicate that feed-forward inhibition is much stronger in the soma than in the dendrites, resulting in a broader integration window in the latter compartment. Thus, the subcellular partitioning of feed-forward inhibition enforces precise coincidence detection in the soma, while allowing dendrites to sum incoming activity over broader time windows.
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              Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics.

              We demonstrate live-cell super-resolution imaging using photoactivated localization microscopy (PALM). The use of photon-tolerant cell lines in combination with the high resolution and molecular sensitivity of PALM permitted us to investigate the nanoscale dynamics within individual adhesion complexes (ACs) in living cells under physiological conditions for as long as 25 min, with half of the time spent collecting the PALM images at spatial resolutions down to approximately 60 nm and frame rates as short as 25 s. We visualized the formation of ACs and measured the fractional gain and loss of individual paxillin molecules as each AC evolved. By allowing observation of a wide variety of nanoscale dynamics, live-cell PALM provides insights into molecular assembly during the initiation, maturation and dissolution of cellular processes.
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                Author and article information

                Contributors
                scimemia@ninds.nih.gov
                m.beato@ucl.ac.uk
                Journal
                Mol Neurobiol
                Molecular Neurobiology
                Humana Press Inc (New York )
                0893-7648
                1559-1182
                22 October 2009
                December 2009
                : 40
                : 3
                : 289-306
                Affiliations
                [1 ]Synaptic Physiology Section, Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, 35 Convent Drive, Building 35, Room 3C-1006, Bethesda, MD 20892-3701 USA
                [2 ]Department of Neuroscience, Physiology and Pharmacology, University College London, London, WC1E 6BT UK
                Article
                8087
                10.1007/s12035-009-8087-7
                2777263
                19844813
                415ce8c0-ec36-4ebf-9bfe-4fa0618afc60
                © The Author(s) 2009
                History
                : 6 July 2009
                : 30 September 2009
                Categories
                Article
                Custom metadata
                © Humana Press Inc. 2009

                Neurosciences
                time course,synapse,diffusion,competitive low-affinity antagonist,neurotransmitter
                Neurosciences
                time course, synapse, diffusion, competitive low-affinity antagonist, neurotransmitter

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