Department of Chemistry and the Beckman Institute, University of Illinois at Urbana-Champaign, 600 S. Mathews Ave., Urbana, Illinois 61801, USA.
AAPS J. 2010 Sep;12(3):443-54. doi: 10.1208/s12248-010-9204-2. Epub 2010 May 19.
To better understand why certain individuals are more vulnerable to cocaine abuse and addiction, we identify peptide markers associated with individual variation in sensitivity to the behavioral effects of cocaine. Previous studies in rats show that low, compared to high, cocaine responders are more sensitive to cocaine-induced behavioral plasticity (sensitization), exhibit enhanced conditioning to cocaine's rewarding effects, and are more motivated to self administer cocaine. In the current study, we combine matrix-assisted laser desorption/ionization mass spectrometry with multivariate statistical methods to analyze tissue extracts from rat dorsal striatum, nucleus accumbens, and medial prefrontal cortex (mPFC) to examine trends in peptide changes that coincide with behavioral phenotype. Peptide profiles of these three regions from individual animals were characterized via mass spectrometry. Resulting mass peaks that were statistically different between these groups were identified using principal component analysis. The mass peaks were then identified in pooled samples via multistage liquid chromatography mass spectrometry. A total of 74 peptides from 28 proteins were sequenced from defined brain regions. Statistically significant changes in peak intensities for seven peptides were found in the mPFC of rats given a single injection of 10 mg/kg cocaine, with low cocaine responders showing approximately 2-fold increase in peak intensities for the acetylated N terminus peptides of stathmin and Hint 1, as well as truncated ATP synthase. These results suggest that distinct peptide profiles in the mPFC are associated with individuals that exhibit reduced sensitivity to the behavioral effects of cocaine.
为了更好地理解为什么某些个体更容易滥用和成瘾可卡因,我们确定了与个体对可卡因行为效应敏感性变化相关的肽标记物。以前在大鼠中的研究表明,与高可卡因反应者相比,低可卡因反应者对可卡因引起的行为可塑性(敏化)更敏感,对可卡因的奖赏效应的条件反射增强,并且更有动力自我给予可卡因。在当前的研究中,我们将基质辅助激光解吸/电离质谱与多变量统计方法相结合,分析大鼠背侧纹状体、伏隔核和内侧前额叶皮质(mPFC)的组织提取物,以检查与行为表型一致的肽变化趋势。通过质谱法对个体动物的这三个区域的肽图谱进行了描述。使用主成分分析确定了这些组之间在统计学上存在差异的质量峰。然后通过多级液相色谱-质谱法在混合样品中鉴定出质量峰。从 28 种蛋白质中总共从定义的脑区中测序了 74 种肽。在给予 10mg/kg 可卡因单次注射的大鼠的 mPFC 中发现了 7 种肽的峰强度的统计学显著变化,低可卡因反应者的 stathmin 和 Hint 1 的乙酰化 N 端肽以及截短的 ATP 合酶的峰强度增加了约 2 倍。这些结果表明,mPFC 中的不同肽谱与对可卡因行为效应敏感性降低的个体有关。