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Journal of Neuroscience, Vol 14, 7521-7528, Copyright © 1994 by Society for Neuroscience
Calcium-activated release of nitric oxide and cellular distribution of nitric oxide-synthesizing neurons in the nervous system of the locust
U Muller and G Bicker
Institut fur Neurobiologie, Berlin, Germany.
Nitric oxide (NO) is generated by a Ca2+/calmodulin-activated NO synthase
and diffuses as a short-lived transcellular messenger through the plasma
membrane. This study investigates the neurochemistry and anatomical
distribution of NO-releasing cells in the CNS of the locust.
Ca2+/calmodulin-activated NO synthase is responsible for fixation-
sensitive NADPH diaphorase (NADPHd) activity in cell homogenates of the
nervous system. Therefore, neurons expressing NO synthase were detected by
NADPHd histochemistry performed in whole-mounts. The anatomical screening
revealed fewer than 1% NADPHd-positive cells in the ventral nerve cord,
some of which were single potentially identifiable neurons, and groups of
cell bodies in several regions of the cerebral ganglion. A prominent
feature of the histochemical survey in the cerebral ganglion is a group of
45 intensely stained cells innervating the olfactory neuropil of the
antennal lobe. A basic requirement for identifying NO as a messenger
molecule is the Ca(2+)-dependent release during nerve cell depolarization.
With a sensitive photometric assay we demonstrated that dissociated cells
from brain areas rich in NADPHd- positive neurons release NO after
stimulation by agents elevating cytoplasmic Ca2+ levels and by the
excitatory neurotransmitter acetylcholine. The combined anatomical and
biochemical experiments therefore provide firm evidence that NO is a
messenger molecule released in the CNS of the locust. Since locust neurons
can be readily grown in primary culture, NO-induced elevations of CGMP
levels and other signal transduction mechanisms in target cells will also
be amenable to a cellular analysis.
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