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The Journal of Neuroscience, May 15, 1998, 18(10):3548-3553
Extrasynaptic Vesicular Transmitter Release from the Somata of
Substantia Nigra Neurons in Rat Midbrain Slices
Erica H.
Jaffe1, 2,
Alain
Marty2,
Albert
Schulte1, and
Robert H.
Chow3
1 Division of Molecular Biology of Neuronal Signaling,
Max-Planck-Institut für Experimentelle Medizin, Göttingen,
Germany D-37075, 2 Arbeitsgruppe Zelluläre
Neurobiologie, Max-Planck-Institut für Biophysikalische Chemie,
Göttingen, Germany D-37077, and 3 Membrane
Biology Group and Department of Physiology, University of Edinburgh
Medical School, Edinburgh, United Kingdom EH8 9AG
Substantia nigra neurons release dopamine from their
somatodendritic regions. A long-unresolved question is whether this
release occurs by exocytosis or by a nonvesicular mechanism. We used
carbon fiber microelectrodes in a brainstem slice to assay secretion from single cell bodies that had been cleared of connective tissue. Amperometry at the carbon fiber microelectrodes revealed unitary events
in ~90% of cells in resting conditions. These events had charge
integrals ranging from a few femtocoulombs to several hundred femtocoulombs (fC). Local glutamate application enhanced the event frequency by 3.5-fold on average and up to 10-fold in highly responsive cells, although the mean charge integral was not modified. Local application of a high K+-containing saline had
effects similar to those of glutamate. The frequency of resting and
stimulated amperometric events was much lower at 21-22°C than at
32-35°C. The addition of Cd2+ (50 µM), a blocker of voltage-dependent
Ca2+ channels, to the bath solution blocked the
stimulatory effects of glutamate. These results suggest that dopamine
is released from the somata of substantia nigra neurons by exocytosis
and that this mechanism is regulated by neuronal electrical activity. More generally, this study demonstrates the applicability of carbon fiber microelectrodes to the measurement of quantal monoamine secretion
in brain slices.
Key words:
substantia nigra; carbon fiber; exocytosis; dopamine; amperometry, pars compacta
Copyright © 1998 Society for Neuroscience 0270-6474/98/18103548-06$05.00/0
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