[Effects of opioids on Ca2+/calmodulin dependent protein kinase signal pathway in NG108-15 cells].
OBJECTIVE
To observe the change of Ca2+/calmodulin dependent protein kinase II (CaMK II) signal pathway in opioid dependent NG108-15 cells.


METHODS
NG108-15 cells were used as an in vitro model system. Competitive protein binding assay and radioimmunoassay were used to examine the intracellular cAMP accumulation. Calmodulin activity was assayed by PDE method. CaMK II activity was assayed by gamma-32 P incorporation of syntide-2.


RESULTS
DPDPE long-term treatment increased calmodulin activity and CaMK II activity in both cytoplasm and nucleus of NG108-15 cells. Specific calmodulin antagonist W-7 was found to significantly inhibit the elevation of calmodulin and CaMK II activity which resulted from DPDPE long-term treatment, and CaMK II inhibitor KN-62 also inhibited elevation of CaMK II activity by DPDPE long-term treatment. When naloxone was added to NG108-15 cells which were long-term treated by DPDPE, calmodulin and CaMK II activity increased, indicating that naloxone withdrawal can increase Ca2+/CaMK II pathway activity.


CONCLUSIONS
The results indicate that Ca2+/CaMK II pathway was involved in the mechanisms of opioids dependence when DPDPE was long-term administered to NG108-15 cells.
Guo
Q M
QM
Department of Pharmacology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China.
Liu
J S
JS
D000701:Analgesics, Opioid / Q000494:pharmacology*; D000818:Animals; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D020881:Enkephalin, D-Penicillamine (2,5)- / Q000494:pharmacology*; D005910:Glioma; D006822:Hybrid Cells; D051379:Mice; D009447:Neuroblastoma; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects*
D000701:Analgesics, Opioid; D002147:Calmodulin; D020881:Enkephalin, D-Penicillamine (2,5)-; D000242:Cyclic AMP; D017871:Calcium-Calmodulin-Dependent Protein Kinases