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1、PROTEOME ANALYSIS REVEALS PROTEIN CANDIDATES INVOLVED IN EARLY STAGES OF BRAIN REGENERATION OF TELEOST FISH蛋白质组分析揭示了再生的硬骨鱼类脑的早期阶段，所涉及的蛋白候选AbstractExploration of the molecular dynamics underlying regeneration in the central nervous system of regeneration-competent organisms has received little attent
2、ion thus far.（在中枢神经系统再生能力的生物再生的分子动力学相关的勘探，迄今很少受到关注）By combining acerebellar lesion paradigm with differential proteome analysis at a post-lesion survival time of 30min, （通过结合一个的小脑病变范例与差异蛋白质组分析后病变的存活时间为30分钟，）we screened for protein candidates involved in the early stages of regeneration in the cerebe
3、llum of such an organism, the teleost fish Apteronotus leptorhynchus. （我们筛选参与再生的早期阶段，在这样一个有机体，硬骨鱼Apteronotus的leptorhynchus小脑的蛋白候选。）Out of 769 protein spots, the intensity of 26 spots was significantly increased by a factor of at least 1.5 in the lesioned hemisphere, relative to the intact hemisphere
4、.（出769个蛋白点，26点的强度明显增加的一个因素至少1.5在损伤半球相对于完整的半球。） The intensity of 9 protein spots was significantly reduced by a factor of at least 1.5.（9蛋白斑点的强度，显着减少了至少为1.5的一个因素）The proteins associated with 15 of the spots were identified by peptide mass fingerprinting and/or tandem mass spectrometry, resulting in t
5、he identification of a total of 11 proteins.（15的斑点与相关的蛋白质，肽质量指纹图谱和/或串联质谱进行鉴定，总共有11种蛋白质的识别导致） Proteins whose abundance was significantly increased include: （蛋白质的丰度显着增加，包括：）erythrocyte membrane protein 4.1N, fibrinogen gamma polypeptide, fructose-biphosphate aldolase C, alpha-internexin neuronal inter
6、mediate filament protein（红细胞膜蛋白4.1N，纤维蛋白原伽玛多肽，果糖二磷酸醛缩酶C，-internexin的神经元中间丝蛋白）, major histocompatibility complex class I heavy chain,（主要组织相容性复合体类重链，） 26S proteasome non-ATPase regulatory subunit 8, （26S蛋白酶体非ATP酶的调节亚基8）tubulin alpha-1C chain,（微管蛋白-1C链） and ubiquitin-specific protease 5.（泛素特异性蛋白酶5。）Pro
7、teins with significantly decreased levels of abundance include（蛋白质水平显着下降丰）: brain glycogen phosphorylase, （脑糖原磷酸化酶）neuron-specific calcium-binding protein hippocalcin,（神经元特异性钙结合蛋白hippocalcin） and spectrin alpha 2. （血影-2）We hypothesize that these proteins are involved in energy metabolism（我们推测，这些蛋白质参
8、与能量代谢）, blood clotting, electron transfer in oxidative reactions, cytoskeleton degradation, apoptotic cell death, synaptic plasticity, axonal regeneration, and promotion of mitotic activity.（血液凝固，电子转移氧化反应，细胞骨架的降解，细胞凋亡，突触可塑性，神经元轴突再生，并促进有丝分裂活动。）Highlights We combine a cerebellar lesion paradigm with d
9、ifferential proteome analysis（我们结合一个的小脑病变范例与差异蛋白质组分析。）. We screen for proteins involved in early stages of regeneration of the cerebellum in a teleost fish. （我们屏幕的蛋白质参与再生的硬骨鱼类的小脑的早期阶段。） We characterize the changes in protein abundance of nearly 800 proteins. 我们近800个蛋白质的蛋白丰度的变化特征。 We identify 11 prot
10、eins and discuss their putative function. 我们确定了11个蛋白，并讨论其假定的功能。 We show that the onset of regeneration occurs within minutes in a regeneration-competent organism. 我们表明，发生再生发病中的再生能力的有机体在几分钟之内。Abbreviations缩略语BVA, biological variance analysis; 生物方差分析;DIA, differential in-gel analysis; （差分凝胶电泳分析）DIGE,
11、difference gel electrophoresis; （差异凝胶电泳）2D-PAGE, two-dimensional polyacrylamide gel electrophoresis; 二维聚丙烯酰胺凝胶电泳;IPG, immobilized pH gradient; 固相pH梯度;MALDI-TOF, matrix assisted LASER desorption/ionization time-of-flight; 基质辅助激光解吸/离子化飞行时间;MHC, major histocompatibility complex; 主要组织相容性复合体MS, mass spec
12、troscopy; 质谱;MS/MS, tandem mass spectrometry; 串联质谱;PMF, peptide mass fingerprinting; 肽质量指纹图谱;SDS, sodium dodecylsulphate十二烷基磺酸钠Key words Apteronotus leptorhynchus; injury; proteomics; 蛋白质组学regeneration; 再生wound healing伤口愈合IntroductionIn most adult vertebrates（在大多数的成年脊椎动物）, and particularly in mammal
13、s, the regenerative capacity of tissue and organs after injury is very limited or absent.（组织和器官损伤后的再生能力是非常有限的或不存在的。） A notable exception is the ability of mammals and other vertebrates to regenerate hepatocytes after liver injury （一个值得注意的例外是哺乳动物和其他脊椎动物的肝损伤后肝细胞的再生能力）(for reviews see Fausto, 2000andMi
14、chalopoulos and DeFrances, 2005). Comparative studies have demonstrated that the regenerative potential is much more pronounced in anamniotes than in amniotes（比较的研究已经表明，再生的潜力在羊膜在anamniotes比更明显）(for review see Stocum, 2006). Well-known examples of the regenerative potential of anamniotes are urodele
15、amphibians, which are capable of regrowing their limbs after amputation （著名的例子的再生潜能anamniotes的的urodele两栖类动物，它们能够重新长出四肢截肢后） (for reviews see Brockes, 1997andNye et al., 2003), and teleost fish, which have retained in adult stages of development not only the capacity to regrow a number of peripheral o
16、rgans, such as fins (for review see Akimenko et al., 2003) or heart （硬骨鱼类，在成人的发展阶段，不仅保留了周边的一些器官，如散热片或心脏的再生能力） (for review see Raya et al., 2004), but also the ability to replace lesioned portions of their central nervous system by generating de novo neurons and glial cells （但也产生从头神经元和神经胶质细胞的能力，以取代其中枢神经系统损伤的部分）(for reviews see Zupanc and Zupanc, 2006a, Becker and Be
