| Opioid addiction has been increasingly considered as a neuropsychiatric disorder and involves in many brain areas, such as hippocampus and cortex. Accumulated evidences have revealed that the neuronal adaptations, resulting in opioid dependence, are related to changes in gene expression and even the structure of the target neurons. However, comprehensive protein profile in brain of animals chronically treated with opioid is still lacking. This study was to apply the proteomic approach to identify proteins significantly changed in opioid dependent rat hippocampus, striatum and cortex. After the rats were treated with ascending doses of morphine to achieve dependence, leading to characteristic withdrawal syndromes, comparative proteomic analysis was performed in the three brain areas of two phases of morphine dependence, long-term exposure to morphine and drug withdrawal. Through analyses of images from two-dimentional gel electrophoresis and statistics, different protein spots distributed mainly in pI 5~8 and molecular masses 17.0~65.0 kDa were determined. After in-gel trypsin digestion, 50 protein spots corresponding to 38 proteins, affected by morphine dependence and withdrawal, were identified by matrix-assisted laser desorption/ ionization-time-of-flight-mass spectrometry combining with nanoelectrospray ionization tandem mass spectrometry (ESI-MS/MS), including 25 proteins that were unknown before to be related with opioid dependence. These proteins are involved in many different cellular and synaptic functions, and some of them are highly related to neuron plasticity. Two proteins were identified as N-terminally acetylated form. Our data pave the way to a better understanding of the underlying neurobiological mechanisms for opioid dependence.
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