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The Journal of Neuroscience, April 1, 2000, 20(7):2523-2533
Highly Localized Ca2+ Accumulation Revealed by
Multiphoton Microscopy in an Identified Motoneuron and Its Modulation
by Dopamine
Peter
Kloppenburg1, 2,
Warren R.
Zipfel2,
Watt W.
Webb2, and
Ronald M.
Harris-Warrick1
1 Department of Neurobiology and Behavior, and
2 Developmental Resource for Biophysical Imaging and
Opto-Electronics, Applied and Engineering Physics, Cornell University,
Ithaca, New York 14853
Calcium is essential for synaptic transmission and the control of
the intrinsic firing properties of neurons; this makes
Ca2+ channels a prime target for neuromodulators. A
combination of multiphoton microscopy and voltage-clamp recording was
used to determine the localization of voltage-dependent
Ca2+ accumulation in the two pyloric dilator (PD)
neurons of the pyloric network in the spiny lobster, Panulirus
interruptus, and its modulation by dopamine. We monitored
[Ca2+]i in fine distal branches in the
neuropil >350 µm below the surface of the ganglion during controlled
voltage steps in voltage clamp. Ca2+ accumulation
originated mostly from small, fairly rare, spatially restricted
varicosities on distal neuritic arborizations. Ca2+
diffused from these point sources into adjacent regions. Varicosities with similar morphology in the PD neuron have been shown previously to
be sites of synaptic contacts. We have demonstrated in earlier studies
that dopamine inhibits activity and greatly reduces synaptic transmission from the PD neuron. In ~60% of the varicosities, the
voltage-activated Ca2+ accumulation was reduced by
exogenous dopamine (DA) (10 4 M). DA
decreased the peak amplitude of Ca2+ accumulation
but had no effect on the rise and decay time. We conclude that DA
reduces chemical synaptic transmission from the PD neurons at least in
part by decreasing Ca2+ entry at neurotransmitter
release sites.
Key words:
calcium; crustacean; central pattern generator; dopamine; motoneuron; multiphoton microscopy; neuromodulation; stomatogastric
ganglion; two-photon microscopy
Copyright © 2000 Society for Neuroscience 0270-6474/00/2072523-11$05.00/0
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