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Volume 17, Number 17,
Issue of September 1, 1997
pp. 6647-6656
Copyright ©1997 Society for Neuroscience
Alteration of Ca2+ Dependence of Neurotransmitter
Release by Disruption of Ca2+ Channel/Syntaxin
Interaction
Received May 9, 1997; revised June 17, 1997; accepted June 20, 1997.
Jens Rettig1,
Christian Heinemann1,
Uri Ashery1,
Zu-Hang Sheng2,
Charles T. Yokoyama2,
William
A. Catterall2, and
Erwin Neher1
1 Department of Membrane Biophysics,
Max-Planck-Institute for Biophysical Chemistry, 37077 Göttingen,
Germany, and 2 Department of Pharmacology, University of
Washington, Seattle, Washington 98195-7280
Presynaptic N-type calcium channels interact with syntaxin and
synaptosome-associated protein of 25 kDa (SNAP-25) through a binding
site in the intracellular loop connecting domains II and III of the
1 subunit. This binding region was loaded into embryonic
spinal neurons of Xenopus by early blastomere injection. After culturing, synaptic transmission of peptide-loaded and control cells was compared by measuring postsynaptic responses under different external Ca2+ concentrations. The relative
transmitter release of injected neurons was reduced by ~25% at
physiological Ca2+ concentration, whereas injection
of the corresponding region of the L-type Ca2+
channel had virtually no effect. When applied to a theoretical model,
these results imply that 70% of the formerly linked vesicles have been
uncoupled after action of the peptide. Our data suggest that severing
the physical interaction between presynaptic calcium channels and
synaptic proteins will not prevent synaptic transmission at this
synapse but will make it less efficient by shifting its Ca2+ dependence to higher values.
Key words:
neuromuscular junction;
Ca2+
dependence;
synaptic transmission;
calcium channel;
synprint;
syntaxin
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