Witzmann Frank A, Li Junyu, Strother Wendy N, McBride William J, Hunter Lawrence, Crabb David W, Lumeng Lawrence, Li Ting-Kai
Department of Cellular and Integrative Physiology, Biotechnology Research and Training Center, Indiana University School of Medicine, 1345 W. 16th Street, Indianapolis, IN 46202, USA.
Proteomics. 2003 Jul;3(7):1335-44. doi: 10.1002/pmic.200300453.
Two-dimensional gel electrophoresis (2-DE) was used to separate protein samples solubilized from the nucleus accumbens and hippocampus of alcohol-naïve, adult, male inbred alcohol-preferring (iP) and alcohol-nonpreferring (iNP) rats. Several protein spots were excised from the gel, destained, digested with trypsin, and analyzed by mass spectrometry. In the hippocampus, 1629 protein spots were matched to the reference pattern, and in the nucleus accumbens, 1390 protein spots were matched. Approximately 70 proteins were identified in both regions. In the hippocampus, only 8 of the 1629 matched protein spots differed in abundance between the iP and iNP rats. In the nucleus accumbens, 32 of the 1390 matched protein spots differed in abundance between the iP and iNP rats. In the hippocampus, the abundances of all 8 proteins were higher in the iNP than iP rat. In the nucleus accumbens, the abundances of 31 of 32 proteins were higher in the iNP than iP rat. In the hippocampus, only 2 of the 8 proteins that differed could be identified, whereas in the nucleus accumbens 21 of the 32 proteins that differed were identified. Higher abundances of cellular retinoic acid-binding protein 1 and a calmodulin-dependent protein kinase (both of which are involved in cellular signaling pathways) were found in both regions of the iNP than iP rat. In the nucleus accumbens, additional differences in the abundances of proteins involved in (i) metabolism (e.g., calpain, parkin, glucokinase, apolipoprotein E, sorbitol dehydrogenase), (ii) cyto-skeletal and intracellular protein transport (e.g., beta-actin), (iii) molecular chaperoning (e.g., grp 78, hsc70, hsc 60, grp75, prohibitin), (iv) cellular signaling pathways (e.g., protein kinase C-binding protein), (v) synaptic function (e.g., complexin I, gamma-enolase, syndapin IIbb), (vi) reduction of oxidative stress (thioredoxin peroxidase), and (vii) growth and differentiation (hippocampal cholinergic neurostimulating peptide) were found. The results of this study indicate that selective breeding for disparate alcohol drinking behaviors produced innate alterations in the expression of several proteins that could influence neuronal function within the nucleus accumbens and hippocampus.
二维凝胶电泳（2-DE）用于分离从未接触过酒精的成年雄性近交系嗜酒（iP）和非嗜酒（iNP）大鼠伏隔核和海马中溶解的蛋白质样品。从凝胶上切下几个蛋白质斑点，脱色，用胰蛋白酶消化，然后进行质谱分析。在海马中，1629个蛋白质斑点与参考图谱匹配，在伏隔核中，1390个蛋白质斑点与参考图谱匹配。两个区域中大约鉴定出70种蛋白质。在海马中，1629个匹配的蛋白质斑点中只有8个在iP和iNP大鼠之间丰度不同。在伏隔核中，1390个匹配的蛋白质斑点中有32个在iP和iNP大鼠之间丰度不同。在海马中，所有8种蛋白质在iNP大鼠中的丰度均高于iP大鼠。在伏隔核中，32种蛋白质中有31种在iNP大鼠中的丰度高于iP大鼠。在海马中，8种差异蛋白质中只有2种可以被鉴定出来，而在伏隔核中，32种差异蛋白质中有21种被鉴定出来。在iNP大鼠的两个区域中均发现细胞视黄酸结合蛋白1和钙调蛋白依赖性蛋白激酶（两者均参与细胞信号通路）的丰度高于iP大鼠。在伏隔核中，还发现了参与以下方面的蛋白质丰度差异：（i）代谢（例如，钙蛋白酶、帕金蛋白、葡萄糖激酶、载脂蛋白E、山梨醇脱氢酶）；（ii）细胞骨架和细胞内蛋白质运输（例如，β-肌动蛋白）；（iii）分子伴侣（例如，grp 78、hsc70、hsc 60、grp75、抑制素）；（iv）细胞信号通路（例如，蛋白激酶C结合蛋白）；（v）突触功能（例如，复合体I、γ-烯醇化酶、syndapin IIbb）；（vi）氧化应激的减轻（硫氧还蛋白过氧化物酶）；以及（vii）生长和分化（海马胆碱能神经刺激肽）。这项研究的结果表明，针对不同饮酒行为的选择性育种在几种蛋白质的表达上产生了先天性改变，这些改变可能会影响伏隔核和海马内的神经元功能。
