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Journal of Neuroscience, Vol 5, 2240-2253, Copyright © 1985 by Society for Neuroscience
Biochemical characterization of a filtered synaptoneurosome preparation from guinea pig cerebral cortex: cyclic adenosine 3':5'-monophosphate- generating systems, receptors, and enzymes
EB Hollingsworth, ET McNeal, JL Burton, RJ Williams, JW Daly and CR Creveling
A particulate preparation was obtained by low speed centrifugation of
guinea pig cerebral cortical homogenates prepared with a Krebs- Henseleit
buffer. Light microscopic examination, using a reflected light differential
interference contrast system, reveals the presence of intact neurons,
axonal fragments, glial cells, and erythrocytes along with an abundance of
small spherical entities (diameter about 1.1 micron) and snowman-shaped
entities (diameter of larger sphere about 1.1 micron, diameter of attached
smaller sphere about 0.6 micron). Many unattached smaller spherical
entities are also present (diameter about 0.6 micron). Pressure filtration
through 5- or 10-micron Millipore filters, followed by low speed
centrifugation and resuspension, removes most of the larger entities to
afford a suspension composed mainly of the small spherical and
snowman-shaped entities. Electron microscopic examination reveals the
presence of many synaptosomes with attached resealed postsynaptic entities.
It is proposed that these correspond to the snowman-shaped entities to be
termed synaptoneurosomes. Accumulations of cyclic AMP elicited by
2-chloroadenosine and histamine, and by combinations of 2-chloroadenosine,
histamine, norepinephrine, and forskolin, are lower in filtered than in
unfiltered preparations, whereas accumulations elicited by forskolin are
unchanged. Levels of adenylate cyclase are reduced by filtration, whereas
levels of phosphodiesterase are unchanged. Filtration reduces levels of
markers for whole cells and endothelial cells, whereas neuronal markers
such as acetylcholinesterase activity and norepinephrine uptake are
increased. Levels of S-100 protein, a marker for glial cells, are not
significantly decreased. There is no apparent change in the density of many
receptors or ion channels. Levels of A1- adenosine and H1-histamine
receptors are increased, whereas levels of so-called peripheral
benzodiazepine-binding sites are decreased.
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