MRC Centre for Synaptic Plasticity University of Bristol
MRC Centre for Synaptic Plasticity, University of Bristol, England NEUROPHYSIOLOGY IN IMAGES Patch clamp Brain Field recording CA 1 CA 2 f. EPSP DG Cytochrome C oxidase stain of a fixed rat barrel cortex f. EPSP 2 m. V Network of cortical neurons EGFP-Sindbis virus infection, CA 1 neuron hippocampus A. Terashima, A. Griffiths & F. Duprat, 2000 Neuron m. V 2 1 f. v. ms Intracellular recordings Patch Clamp Acute brain slices Adapted from Agmon & Connors 1991, Neurosci. 41, pages 365 -379 100 200 Stimulus duration (ms) Adapted from E. J. Coan, Ph. D thesis 1988, UOB 10° From Feldmeyer et al. 1999, J. Physiol. 521, page 171 ms 100 200 Stimulus duration (ms) Adapted from C. H. Davies, Ph. D thesis 1991, UOB From Agmon & Connors 1991, Neurosci. 41, page 370 4 p. s. Dendritic extracellular recording Subthreshold Suprathreshold 55° 8 m. V Stimulation Subthreshold Suprathreshold 10 ms Overnight slice culture 100 200 Stimulus duration (ms) Preparations Blind Patch Clamp Neurons in culture EPSC 50 p. A Neuronal Network 10 ms Thalamocortical slices 7 days long term culture. Injection in CA 3 pyramidal neurons Courtesy of Akira Terashima, 1999 2 12 4 m. V From G. Paxinos & C. Watson, The Rat Brain, 1998 Hippocampal slices 4 Somatic extracellular recording CA 3 From H. ELLIS, B. LOGAN & A. DIXON “Human cross-sectional anatomy" 1991, Ed. Butterworth Heinemann Extracellular recording EPSP slope (m. V/ms) Long term slice culture Population spike (m. V) Nervous system Fiber volley (m. V) Dr Fabrice DUPRAT, March 2000 50 ms High resolution Patch Clamp Hippocampal neuron in culture (P 10 -14) Courtesy of Lisa Pickard, 1999 Imaging Electrophysiology Localisation Functional properties 5 mm Molecular Biology / Biochemistry Molecular properties Axon Synaptic bouton in rat CA 1 stratum radiatum Electron microscopy. From Shepherd et al. 1998, J. Neuroscience 18, p. 8302 Axon Rat CA 3 Pyramidal cell 3 D reconstruction of multiple spine boutons From Toni et al. 1999, Nature 402, p. 423 Dendrite Cl- GABA, Isoguvacine, Muscimol Bicuculline, SR 95531 GABAB R 1 a, b, c, d R 2 Benzodiazepines, Flumazenil, ZK 93426 Glu. R 1, 2, 3, 4 Glu. R 5, 6, 7 Ca 2+ impermeable Low affinity, Ca 2+ impermeable Edited Glu. R 2 Edited Glu. R 5, 6 3 D representation of the torpedo nicotinic acetylcholine receptor Ca 2+ K+, Na+ L-Baclofen, GABA Phaclofen CGP 35348, 2 -OH saclofen, CGP 55845 Competitive Modulatory NMDA, L-Glutamate, L-Aspartate, Ibotenate Glycine, Serine D-a. AA D-AP 5, CGS 19755, CGP 37849, CGP 39551, AP 7, CPP Na+, K+ (Ca 2+, Mg 2+) Na+, K+ (Ca 2+) Quisqualate, AMPA, Domoate, Glutamate, Kainate, Glutamate, Domoate, AMPA, ATPA 7 -chloro GYKI 53655*, kynurenate, GYKI 52466*, CNQX, NBQX, + 2+ NO, H , Mg CNQX DNA m. Glu 2, 3 m. Glu 4, 6, 7, 8 LY 382884, LY 294486, NS 102, NBQX, CNQX Gq/11 PLC Gi/o AC Quisqualate, LY 379268, (S)-DHPG, L-CCG-1, Glutamate, DCG-IV, ACPD, (2 R, 4 R)-APDC, Ibotenate, LY 354740 , CHPG 5 Glutamate, (1 S, 3 S)-ACPD L-AP 4 , L-SOP, Glutamate, L-CCG-1 AIDA, MCCG, 1 LY 367385 , LY 341495, MPEP 5*, PCCG-IV 1 CPCCOEt * MCPG LY 341495, MAP 4, CPPG x 4= (x 5 ? ) Inter-m. EPSC Interval (s) 0 20 Time (min) 40 60 Long Term Depression: LTD m. EPSC Amplitude (p. A) From Noel et al. Neuron 23, p. 2 2) Na+-channel dependent: spontaneous = spontaneous action potential (no TTX) s. EPSP, s. EPSC, s. IPSP, s. IPSC (* non competitive, 1, 5 m. Glu 1 or 5 only) = activity of N synapses EPSP, EPSC, IPSP, IPSC -20 -80 10 Time (min) +30 0 -30 Frequency (Hz) 20 From Bear 1996 Proc. Natl. Acad. Sci. USA 93, p. 13455 25 15 20 ms 5 -80 -40 0 EPSC amplitude (p. A) Depression of EPSCs by pep 2 m From Lüthi et al. 1999 Neuron 24, p. 392 F. Aguila & F. Duprat, 1998 100 -20 Evoked activity Glu. Rx Axon m. Glu 1, 5 KA 1, 2 F. Duprat, march 2000 From Green et al. 1998, Neuron 20, p. 427 -444 Synaptic plasticity High affinity Gi/o AC Flunitrazepam, Zolpidem, Abecarnil NR 1 a, b, c, d, e, NR 2 A, B, C, D Low affinity, Ca 2+ permeable EPSC Amplitude (%) Subunits g 1, 2, 3 d 1 r 1, 2, 3 e 1 Ca 2+ permeable III Change in EPSPslope (%) Varicosity Dendrite a: 6 s/u, b: 4 s/u I Types: II Long Term Potentiation: LTP EPSC Amplitude (%) Mitochondria Vesicles Postsynaptic spine BZ m. EPSP, m. EPSC, m. IPSP, m. IPSC 5 p. A Glial cell GABA Signals PSD Presynaptic bouton Agonists Axon NMDA Sites Antagonists Synapse Spine GABAB Metabotropic Glutamate Receptors Non NMDA AMPA Kainate Structures Dendrite GABAA 1) Na+-channel independent: minis = spontaneous neurotransmitter release (TTX) Normalised frequency Rat CA 1 Pyramidal cell labeled with EGFP Two photon laser scanning microscopy From Maletic-Savatic et al. 1999, Science 283, p. 1924 GABA Receptors Names Synapses Ionotropic Glutamate Receptors Spontaneous activities Cumulative probability Surface and total distribution of AMPA receptors in cultured hippocampal Neurones. Noel et al. 1999, Neuron 23 p. 365 -376 Dendritic tree Receptors Glu. R 2 EPSC amplitude (p. A) Glu. R 1 -4 100 -20 0 20 Time (min) 40 60
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