Journal of Neuroscience, Vol 14, 2402-2407, Copyright © 1994 by Society for Neuroscience
Direct observation of the effect of autoreceptors on stimulated release of catecholamines from adrenal cells
R Zhou, G Luo and AG Ewing
Department of Chemistry, Pennsylvania State University, University Park 16802.
The direct effect of alpha 2-autoreceptors was studied by measuring the
effects of piperoxan, an alpha 2-autoreceptor antagonist, and clonidine, an
agonist on catecholamine exocytosis, from single bovine chromaffin cells in
culture. Catecholamine release was elicited by stimulation with 100 microM
nicotine and was monitored electrochemically with a carbon-fiber
microelectrode placed adjacent to the cell. These electrodes allowed the
number of exocytotic release events to be monitored and reported as total
charge for release following a specific stimulus. Repeated stimulation with
100 microM nicotine showed that total release caused by the second exposure
to nicotine was 32% of the first, and release caused by the third exposure
to nicotine was 80% of the second. Total release of catecholamine increased
significantly after application of 20 microM piperoxan relative to a
control application of balanced salt solution. Application of 20 microM
piperoxan alone did not cause release. After the cells were incubated in
culture medium containing 20 microM clonidine, a significant decrease in
nicotine-stimulated catecholamine release was observed. These results
confirm that there are autoreceptors on chromaffin cells and, when
relatively high levels of catecholamine are released, the catecholamine
stimulates the alpha 2- autoreceptors, which inhibits subsequent release
through a negative feedback mechanism. In addition to piperoxan, the
sympathomimetic drug amphetamine also increases quantal release after
application of nicotine. Amphetamine increases the extracellular
concentration of catecholamine, and these data appear to indicate that at
least part of the pharmacology of amphetamine might involve blocking
catecholamine autoreceptors.