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Vol. 283, Issue 3, 1445-1452, 1997
Department of Pharmacology, The University of Michigan Medical
School, Ann Arbor, Michigan
Repeated, intermittent treatment of rats with amphetamine followed by a
withdrawal period leads to an enhancement in amphetamine-induced dopamine release. We previously reported an increased stoichiometry of
site 3-phospho-synapsin I and increased levels of
phospho-Ser41-neuromodulin in striatum after repeated
amphetamine. In this study, we examined whether the enhanced
amphetamine-induced dopamine release and increased levels of these
phosphoproteins would be detected in synaptosomes from rats pretreated
and withdrawn from repeated amphetamine. Enhanced amphetamine-induced
dopamine release was detected in striatal synaptosomes from rats
treated with repeated amphetamine compared with controls. The enhanced
dopamine release was Ca++ dependent. State-specific
antibodies were used to measure the levels of site 3-phospho-synapsin
I, phosphorylated by CaM kinase II, and
phospho-Ser41-neuromodulin, phosphorylated by protein
kinase C, in incubated striatal S1 fractions and synaptosomes. The
levels of site 3-phospho-synapsin I and
phospho-Ser41-neuromodulin were increased by 40% and 30%,
respectively, in amphetamine-pretreated rats compared with controls.
Total neuromodulin and synapsin I was not altered. There was a
significant 26% increase in CaM kinase II activity in the synaptosomes
from amphetamine-pretreated rats but no change in content. No change in
protein kinase C activity or content of the 
-isozyme was detected
after repeated amphetamine. Our results demonstrate that the enhanced
amphetamine-induced dopamine release and occurring after repeated
amphetamine can be detected in synaptosome preparations. Repeated
amphetamine leads to alterations in phosphorylation/dephosphorylation
activities that can be detected in the incubated synaptosomes. Because
the enhanced amphetamine-induced dopamine release after repeated
amphetamine appears to be Ca++ sensitive, it is possible
that the altered phosphorylation systems, and perhaps site
3-phospho-synapsin I and phospho-Ser41-neuromodulin, play a
role in the enhanced dopamine release.
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