药物成瘾的蛋白质组学研究

药物成瘾涉及脑内多种蛋白含量、结构及功能的复杂改变,主要涉及代谢酶类、细胞骨架蛋白、分子伴侣、细胞内信号途径相关蛋白、突触功能相关蛋白和氧化还原相关蛋白等类型。蛋白质组学能对生理与病理状态下的体液、组织或细胞中基因组编码的所有蛋白质组分进行高通量的综合分析,针对筛选出的有意义“候选”蛋白（candidate protein）进行深入验证研究,不仅可能从蛋白质水平上阐明成瘾的神经生物学作用机制,还有助于建立诊断标准,发现抗成瘾药物治疗的潜在靶点。     

植物微丝骨架的研究进展

微丝骨架是细胞骨架的重要组成部分,在各种细胞活动中都发挥着重要作用。微丝骨架的主要组成部分是肌动蛋白和肌动蛋白结合蛋白,参与细胞形态建成、物质运输和信号转导等生命活动。通过鬼笔环肽标记或表达荧光融合蛋白等方法,国内外许多学者对植物微丝骨架的组成、功能等进行了大量的研究,并取得了一些成果。基于前人的研究,本研究从组成、功能及研究方法三个方面对植物微丝骨架的进行概述。     

淋巴细胞骨架的研究

淋巴细胞骨架的研究陈吉龙，王平（中国科学院发育生物学研究所北京１０００８０）（北京大学生物系）关键词淋巴细胞，细胞骨架，核骨架细胞骨架（包括细胞质骨架和核骨架）的研究，加深了人们对细胞结构和功能的认识。淋巴细胞是形态和功能均较多变的一类细胞，有关其细...     

原生动物的细胞骨架蛋白及其功能组件

目前在原生动物中发现了许多新的细胞骨架蛋白,如中心元蛋白、副鞭毛杆蛋白等。深入研究发现,原生动物的细胞骨架在细胞的模式形成,细胞核的遗传中也具有重要作用。从功能组件角度着眼研究细胞骨架的功能,将有助于了解细胞骨架的进化机制。     

细胞运动、细胞迁移与细胞骨架研究进展

细胞定向运动与细胞骨架的动态循环密切相关。运动细胞在其伪足前沿依靠细胞骨架的不断聚合推动细胞膜的前进,在基部靠近细胞体部位通过细胞骨架的不断解聚收缩拖拉细胞体向前运动,细胞骨架的聚合与解聚通过伪足与支撑表面的吸附与解吸附而偶连。肌动蛋白组成的微丝骨架是大多数运动细胞的主要成分。外界刺激引起微丝细胞骨架动态变化的信号通路已逐步明了。线虫精子细胞的运动行为与阿米巴变形运动相似,但是在线虫精子细胞中没有肌动蛋白,而是以精子主要蛋白为基础形成细胞骨架驱动精子细胞的运动。与肌动蛋白不同,精子主要蛋白没有分子极性、ATP结合位点和马达蛋白。通过比较研究以上两种运动体系将有助于在分子水平上进一步阐明细胞运动的机理。     

Sindbis病毒的蛋白质合成与细胞骨架的关系

我们以Sindbis病毒感染BHK-21细胞为模式,研究了病毒的感染与细胞骨架的关系。结果显示:在病毒感染早期,细胞的蛋白质合成迅速被抑制,细胞的多聚核糖体(polysome)和mRNA从骨架上脱落,而病毒的RNA结合到骨架上。我们的结果还进一步表明,病毒的RNA是通过其3′-尾端与骨架结合的。另一方面在对Sindbis病毒非结构蛋白在体内与体外合成与加工的比较中,我们发现病毒蛋白在体外翻译加工的速度远低于体内,并且出现很多未成熟蛋白(premature protein),这种区别可能在某种程度上反应细胞骨架在蛋白质合成与加工中的作用。此外,在用秋水仙素和细胞松驰素B破坏微管和微丝后,病毒非结构蛋白的合成与加工没有明显变化,而结构蛋白的合成则受到明显的抑制。这表明病毒的两类蛋白的合成所依赖的细胞骨架成分可能有所不同,在结构蛋白合成过程中,微丝和微管起了重要作用,在非结构蛋白合成过程中,中间丝很可能起了重要作用。     

细胞的骨架系统

细胞骨架是一类复杂的蛋白质纤维结构,广泛地存在于动物细胞、植物细胞甚至一些原生动物与酵母中。细胞骨架按分布区域可分为胞质骨架和细胞核骨架,胞质骨架又具有三种类型:微管、微丝和中等纤维.胞质骨架和核骨架以及三种胞质骨架之间的结构、性质和功能上是有所区别的,但另一方面它们又协调地参予细胞的一系列生理活动,共同组成了细胞的骨架系统。六十年代初,波特(K·Porter)等第一次用电镜证明了细胞质中骨架结构的多样性,他们发现几乎每一个真核细胞的胞质中都存在三种类型的骨架结构,即微管、微丝和中等纤维。之后,对它们的结构、性质和功能进行了深入的研究。七十年代以来,在细胞核中又发现了一个形态类似于胞质骨架、蛋白质性质的网架结构——细胞核骨架(简称核骨架)对它可能的作用也有了初步的认识,这些发现丰富了骨架系统的内容。现在,已经证实胞质骨架和核骨架在结构与功能上是密切联系的,两者构成了统一的细胞骨架体系,对细胞生长、运动及细胞分化等过程起着重要的作用。     

植物微管和微丝骨架研究的进展

从以上叙述的资料中可以看出,近年来在植物微管蛋白的分离及其化学性质、微管的组织中心、微管的异质性、微丝的分布,以及微管和微丝骨架的功能及基因调节等方面的研究取得不少新的进展;特别是从植物中直接分离微管蛋白取得成功、以及微管蛋白异型、微管冷稳定性与植物抗寒性的关系及微丝分布广泛性等的发现,对植物细胞骨架的进一步研究具有重要意义。     

SM22α在细胞骨架组构及血管重塑中的作用

血管平滑肌细胞（VSMC）表型转化是动脉粥样硬化、高血压和血管成形术后再狭窄等血管重塑性疾病的共同病理生理过程。VSMC表型转化过程中平滑肌特异基因的表达变化和细胞骨架的组构是当前研究的热点问题之一。平滑肌22α（SM22α）是近年发现的一种VSMC分化标志物,其表达具有平滑肌组织特异性和细胞表型特异性,该蛋白作为一种肌动蛋白细胞骨架相关蛋白参与VSMC骨架组构和收缩调节。本文就SM22α的结构特征及其在VSMC骨架组构和血管重塑中的作用机制进行综述。     

微管骨架在植物适应低温胁迫中的功能研究进展

植物细胞骨架对低温胁迫的响应是近年来研究的一个活跃的前沿领域。本文综述了该领域研究的进展情况和发展趋势:植物微管骨架的结构和功能的简介,低温诱导植物细胞微管骨架稳定性的变化;并对微管骨架在冷信号传导中的作用进行了探讨。     

Proteomics Analysis of Dorsal Striatum Reveals Changes in Synaptosomal Proteins following Methamphetamine Self-Administration in Rats

Methamphetamine is a widely abused, highly addictive drug. Regulation of synaptic proteins within the brain’s reward pathway modulates addiction behaviours, the progression of drug addiction and long-term changes in brain structure and function that result from drug use. Therefore, using large scale proteomics studies we aim to identify global protein expression changes within the dorsal striatum, a key brain region involved in the modulation of addiction. We performed LC-MS/MS analyses on rat striatal synaptosomes following 30 days of methamphetamine self-administration (2 hours/day) and 14 days abstinence. We identified a total of 84 differentially-expressed proteins with known roles in neuroprotection, neuroplasticity, cell cytoskeleton, energy regulation and synaptic vesicles. We identify significant expression changes in stress-induced phosphoprotein and tubulin polymerisation-promoting protein, which have not previously been associated with addiction. In addition, we confirm the role of amphiphysin and phosphatidylethanolamine binding protein in addiction. This approach has provided new insight into the effects of methamphetamine self-administration on synaptic protein expression in a key brain region associated with addiction, showing a large set of differentially-expressed proteins that persist into abstinence. The mass spectrometry proteomics data are available via ProteomeXchange with identifier PXD001443.     

甲基苯丙胺成瘾的表观遗传学机制

苯丙胺类兴奋剂是全世界第二大滥用程度的药物,甲基苯丙胺作为苯胺类兴奋剂中的主要药物,是中国滥用的“头号毒品”。而现有的研究对甲基苯丙胺成瘾机制尚不清晰,且临床上对药物成瘾的治疗依然存在无药可医的局面。因此,发现新的成瘾机制和治疗策略尤为迫切。甲基苯丙胺成瘾与额前叶皮质（mPFC）、中脑腹侧被盖区（VTA）和伏隔核（NAc）中的多巴胺（DA）、谷氨酸（Glu）、去甲肾上腺素（NE）和血清素（SNRIS）等神经递质的异常释放有关。研究表明,这些神经递质受到表观遗传机制中组蛋白乙酰化、甲基化、泛素化和非编码RNA等调节,某些基因的表达在甲基苯丙胺的诱导过程中增强或被抑制,导致甲基苯丙胺依赖性产生。本文将针对表观遗传学对甲基苯丙胺成瘾机制的影响进行着重论述,以期推进临床开发甲基苯丙胺戒断药物的研究。     

miRNAs与药物成瘾

药物成瘾是一种慢性复发性脑病,主要表现为不可控制的对药物持续渴求和戒断后的高复吸。目前观点认为,成瘾是中脑腹侧被盖（ventral tegmental area,VTA）到伏隔核（nucleus accumbens,NAc）脑区多巴胺能奖赏通路中神经可塑性发生改变而导致的一种神经精神疾病。基因表达变化在神经可塑性中发挥着重要作用,但成瘾药物导致相关脑区结构和功能改变的机制还不甚清楚。微小RNAs（microRNAs,miRNAs）是一类非编码RNA,主要通过结合靶基因mRNA 3′非翻译区（3′untranslated region,3′UTR）,在转录后水平阻断其翻译成蛋白质或触发其不稳定而降解。越来越多的研究证实,miRNAs参与调节成瘾相关神经可塑性的变化。本文较系统地阐述miRNAs在药物成瘾中的作用研究进展,将为深入阐明药物成瘾的机制以及药物成瘾临床有效干预和诊治提供新思路。     

The Role of the Axonal Cytoskeleton in Diabetic Neuropathy

The neuropathy associated with diabetes includes well documented impairment of axonal transport, a reduction in axon calibre and a reduced capacity for nerve regeneration. All of those aspects of nerve function rely on the integrity of the axonal cytoskeleton. Alterations in the axonal cytoskeleton in experimental diabetes include an insulin-dependent non-enzymatic glycation of actin that is reflected in increased glycation of platelet actin in the clinical situation. There is a reduced synthesis of mRNA for the isoforms of tubulin that are associated with nerve growth and regeneration and an elevated non-enzymatic glycation of peripheral nerve tubulin in both diabetic patients and diabetic animals. mRNAs for neurofilament proteins are selectively reduced in the diabetic rat and the post-translational modification of at least one of the neurofilament proteins is altered. There is some evidence that altered expression of isoforms of protein kinases may contribute to these changes.     

Changes in Organization and Axonal Transport of Cytoskeletal Proteins During Regeneration

Changes in solubility and transport rate of cytoskeletal proteins during regeneration were studied in the motor fibers of the rat sciatic nerve. Nerves were injured by freezing at the midthigh level either 1-2 weeks before (experiment I) or 1 week after radioactive labeling of the spinal cord with L-[35S]methionine (experiment II). Labeled proteins in 6-mm consecutive segments of the nerve 2 weeks after labeling were analyzed following fractionation into soluble and insoluble populations with 1% Triton at 4 degrees C. When axonal transport of newly synthesized cytoskeleton was examined in the regenerating nerve in experiment I, a new faster component enriched in soluble tubulin and actin was observed that was not present in the control nerve. The rate of the slower main component containing most of the insoluble tubulin and actin together with neurofilament proteins was not affected. A smaller but significant peak of radioactivity enriched in soluble tubulin and actin was also detected ahead of the main peak when the response of the preexisting cytoskeleton was examined in experiment II. It is thus concluded that during regeneration changes in the organization take place in both the newly synthesized and the preexisting axonal cytoskeleton, resulting in a selective acceleration in rate of transport of soluble tubulin and actin.     

Drug addiction: the neurobiology of behaviour gone awry

Drug addiction manifests as a compulsive drive to take a drug despite serious adverse consequences. This aberrant behaviour has traditionally been viewed as bad "choices" that are made voluntarily by the addict. However, recent studies have shown that repeated drug use leads to long-lasting changes in the brain that undermine voluntary control. This, combined with new knowledge of how environmental, genetic and developmental factors contribute to addiction, should bring about changes in our approach to the prevention and treatment of addiction.     

The potential biomarkers of drug addiction: proteomic and metabolomics challenges

Drug addiction places a significant burden on society and individuals. Proteomics and metabolomics approaches pave the road for searching potential biomarkers to assist the diagnosis and treatment. This review summarized putative drug addiction-related biomarkers in proteomics and metabolomics studies and discussed challenges and prospects in future studies. Alterations of several hundred proteins and metabolites were reported when exposure to abused drug, which enriched in energy metabolism, oxidative stress response, protein modification and degradation, synaptic function and neurotrasmission, etc. Hsp70, peroxiredoxin-6 and α- and β-synuclein, as well as n-methylserotonin and purine metabolites, were promising as potential biomarker for drug addiction.     

Genes and Pathways Co-associated with the Exposure to Multiple Drugs of Abuse,Including Alcohol,Amphetamine/Methamphetamine,Cocaine, Marijuana,Morphine, and/or Nicotine: a Review of Proteomics Analyses

Drug addiction is a chronic neuronal disease. In recent years, proteomics technology has been widely used to assess the protein expression in the brain tissues of both animals and humans exposed to addictive drugs. Through this approach, a large number of proteins potentially involved in the etiology of drug addictions have been identified, which provide a valuable resource to study protein function, biochemical pathways, and networks related to the molecular mechanisms underlying drug dependence. In this article, we summarize the recent application of proteomics to profiling protein expression patterns in animal or human brain tissues after the administration of alcohol, amphetamine/methamphetamine, cocaine, marijuana, morphine/heroin/butorphanol, or nicotine. From available reports, we compiled a list of 497 proteins associated with exposure to one or more addictive drugs, with 160 being related to exposure to at least two abused drugs. A number of biochemical pathways and biological processes appear to be enriched among these proteins, including synaptic transmission and signaling pathways related to neuronal functions. The data included in this work provide a summary and extension of the proteomics studies on drug addiction. Furthermore, the proteins and biological processes highlighted here may provide valuable insight into the cellular activities and biological processes in neurons in the development of drug addiction.     

Neurobiological Signatures of Alcohol Dependence Revealed by Protein Profiling

Alcohol abuse causes dramatic neuroadaptations in the brain, which contribute to tolerance, dependence, and behavioral modifications. Previous proteomic studies in human alcoholics and animal models have identified candidate alcoholism-related proteins. However, recent evidences suggest that alcohol dependence is caused by changes in co-regulation that are invisible to single protein-based analysis. Here, we analyze global proteomics data to integrate differential expression, co-expression networks, and gene annotations to unveil key neurobiological rearrangements associated with the transition to alcohol dependence modeled by a Chronic Intermittent Ethanol (CIE), two-bottle choice (2BC) paradigm. We analyzed cerebral cortices (CTX) and midbrains (MB) from male C57BL/6J mice subjected to a CIE, 2BC paradigm, which induces heavy drinking and represents one of the best available animal models for alcohol dependence and relapse drinking. CIE induced significant changes in protein levels in dependent mice compared with their non-dependent controls. Multiple protein isoforms showed region-specific differential regulation as a result of post-translational modifications. Our integrative analysis identified modules of co-expressed proteins that were highly correlated with CIE treatment. We found that modules most related to the effects of CIE treatment coordinate molecular imbalances in endocytic- and energy-related pathways, with specific proteins involved, such as dynamin-1. The qRT-PCR experiments validated both differential and co-expression analyses, and the correspondence among our data and previous genomic and proteomic studies in humans and rodents substantiates our findings. The changes identified above may play a key role in the escalation of ethanol consumption associated with dependence. Our approach to alcohol addiction will advance knowledge of brain remodeling mechanisms and adaptive changes in response to drug abuse, contribute to understanding of organizational principles of CTX and MB proteomes, and define potential new molecular targets for treating alcohol addiction. The integrative analysis employed here highlight the advantages of systems approaches in studying the neurobiology of alcohol addiction.