scuola di neuroscienze · dip. neuroscienze, torino valentina carabelli andrea marcantoni valentina...
TRANSCRIPT
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Scuola di NeuroscienzeUniversità degli Studi di Torino
Corso di DottoratoTorino, 7 Settembre 2006
Canali del calcio presinaptici: distribuzione e neurotrasmissione
CNR-INFM
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Presynaptic Ca2+ channels:types and distribution
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from Purves et al., 2004 Neuroscience, Sinauer Associated Inc., 3rd edition
The synaptic transmission
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The classical approach to identify the contribution of presynaptic Ca2+ channels
in central synapses
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Baldelli et al. (2002) Europ J Neurosci 16: 2297
prepost
Action potential-evoked GABAergic IPSCs
Monosynaptic preparation
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Ca2+ entry at cerebellar excitatory synapses is
controlled by N, P/Q and R-type channels
Mintz et al., 1995 Neuron 15: 675
presynaptic Ca2+-entry(fluorescence)
presynaptic stimuli
postsynapticrecording
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Mintz et al., 1995 Neuron 15: 675
…. while neurotransmitter release is controlled only by
N and P/Q-type channels
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GABA release is controlled by N- and P/Q-type Ca channels in interneurons of hippocampal slices
N-type channels
P/Q-type channels
N and P/Q-type channels
Wilson et al., 2001 Neuron 31: 453
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From these experiments we can conclude that:
• Ca2+ enters through various voltage-gated Ca2+ channels (N, P/Q, R ….. L, T)
• Secretion is strictly associated to N and P/Q-type channels, which are possibly co-localized to the release sites
• There are release sites controlled by P/Q, others controlled by N and a group controlled by N and P/Q
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Reid et al., 2003 TINS 26: 683
“Supra-additivity” supports a model of synaptic terminals with a non-uniform distribution of Ca2+
channels
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A direct approach to study presynaptic Ca2+ channels and neurosecretion:
the giant synapse of the calyx of Held
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The calyx of Held allows to record directly presynaptic and postsynaptic currents
principal neuron in the MNTB
presynapticcalyx
0.3 µmnucleus
Saetzler et al., 2002 J. Neurosci., 22: 10567
pre
post
MNTB (Medial Nucleus of the Trapezoid Body) in the brainstem of the acustic pathway
Glutamatergic giant synapse
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aga aga
P-type channels control fully the EPSCs but partially presynaptic [Ca2+]
GVIA
GVIA
N-type channels control partially the EPSCs and presynaptic [Ca2+]
Wu et al., 1999 J Neurosci, 19: 726
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P-type channels have a steeper Ca2+-dependence than N-type channels
P-type N-type
Wu et al., 1999 J Neurosci, 19: 726
Release [Caterminal]m
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P/Q-type and N-type channels co-localize with synaptotagmin
synaptotagminCa2+ channels merge
P/Q-type (α1A)
N-type (α1B)
R-type (α1E)
Wu et al., 1999 J Neurosci, 19: 726
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Why N and P/Q channels are co-localized at the release sites?
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Catteral, 2000 Annu Rev Cell Dev Bio. 16: 521
Exist a strict interaction between Ca2+ channels (P/Q and N), membrane-binding and vesicle-binding proteins
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from Purves et al., 2004 Neuroscience, Sinauer Associated Inc.
Co-localization of N and P/Q type channels at the release sites is favoured by the interaction of a
channel region (synprint) with syntaxin and other vesicle binding proteins
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Spafford & Zamponi, 2003 Curr Op Neurobiol 13: 308
The “synprint region”
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Are N and P/Q-type channels the only voltage-gated Ca2+ channels controlling
neurotransmitter release?
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Murakami et al., 2002 Brain Research, 951: 121
In isolated hippocampal neurones, focal stimulation is controlled by L-type channels
IPSCs
In slices, presynaptic response is controlled by N and P/Q-type channels
IPSCs
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……. a possible explanation
Murakami et al., 2002 Brain Research, 951: 121
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Jensen et al., 1999 J. Neurophysiol 81: 1225
In isolated hippocampal neurons the post-tetanic potentiation (PTP) iscontrolled by L-type channels
IPSCs (before PTP,at normal
stimulation, 0.2 Hz)
IPSCs (after PTP,at normal
stimulation, 0.2 Hz)
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R- and L-type Ca2+ channels contribute to secretion during prolonged presynaptic stimuli
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Baldelli et al. (2002) Europ J Neurosci 16: 2297
prepost
Action potential-evoked GABAergic IPSCs
Monosynaptic preparation
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Contribution of N and P/Q-type channels to AP-evoked IPSCs in cultured hippocampal neurons
Baldelli et al. (2005) J Neurosci 25: 3358
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KCl-evoked IPSCs induce long lasting eIPSCs with comparable amplitudes and pharmacology
Baldelli et al. (2002) Europ J Neurosci 16: 2297
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Contribution of R-type channels is enhanced during KCl-evoked IPSCs
Baldelli et al. (2005) J Neurosci 25: 3358Baldelli et al. (2002) Europ J Neurosci 16: 2297
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Conclusions
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Vesicle docking, priming, depletion and replenishment are all Ca2+-dependent processes
P/Q P/QNN
N
…….. are they controlled by the same Ca channel types?
R
LL
L
R
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Dip. Neuroscienze, TorinoValentina Carabelli Andrea MarcantoniValentina ComunanzaDaniela Guido
Pietro Baldelli (Dip. Biomedicina Sperimentale, Genova)Monica Novara (Novartis, Milano)Tiziana Cesetti (Dept Neuroscience, Heidelberg)Jesus-Miguel Hernandez-Guijo (Dep. Farmacologia, UAM, Madrid)Anna Giancippoli (Novartis, Milano)Paolo Cesare (E.B.R.I., Roma)Valeria Magnelli (Dip. Scienze e Tecnologie Avanzate, Alessandria)Michele Lovallo (C.N.R., Università di Potenza)Daniela Platano (Dip. Fisiologia Umana e Generale, Bologna)
Emanuele Sher (Ely Lilly Res. Center, Surrey, UK)Giorgio Aicardi (Dip. Fisiologia Umana e Generale, Bologna)Claudio Grassi (Dip. Fisiologia Umana, Univ. Cattolica del Sacro Cuore, Roma)Marcello D’Ascenzo
Antonio Garcia (Dep. Farmacologia, UAM, Madrid)Almudena AlbillosLuis GandiaCarmen Montiel
Antonio Artalejo (Dep. Toxicologia y Farmacologia, Univ Complutense, Madrid)
Helmut Zucker (M.P.I. fur Neurobiologie, Martinsried, Germany)Dieter Swandulla ( Dept. Neuropharmacology, Bonn, Germany)