孤独症与MeCP2基因相关性研究进展CSCD

研究表明甲基化Cp G结合蛋白2(methyl Cp G binding protein 2,Me CP2)基因的突变和功能缺失会导致神经系统障碍,有些孤独症患儿中存在Me CP2基因编码区突变。本文根据国内外阐述Me CP2与神经系统疾病的文献,梳理孤独症患儿与Me CP2基因的相关性,探讨Me CP2基因对孤独症发病的影响。     

Rett综合征相关基因MeCP2敲除大鼠模型的构建及分析

MeCP2(Methyl CpG binding protein 2)基因突变可导致Rett综合征(Rett syndrome, RTT)。目前已报道的MeCP2敲除小鼠表型与RTT病人症状存在显著差异。为探索MeCP2在脑发育中的作用及其导致RTT的机制,本研究利用CRISPR/Cas9技术构建了MeCP2基因敲除大鼠模型。通过构建靶向敲除MeCP2基因的载体,体外将Cas9 mRNA和sgRNA显微注射到SD大鼠受精卵中,在MeCP2基因exon2中造成移码突变,从而获得MeCP2基因敲除大鼠。利用测序和Western blotting方法鉴定MeCP2敲除大鼠,并对其表型和行为学特征进行分析,发现MeCP2敲除大鼠体重降低,存在焦虑倾向和认知缺陷。本研究成功构建了MeCP2基因敲除大鼠模型,其表型类似人类RTT患者的症状,为后续MeCP2功能研究提供了更好的动物模型。     

快速老化模型小鼠SAMP8行为学的增龄性变化

目的：对快速老化模型小白鼠SAMP8老化征象和学习记忆能力的增龄性变化进行系统研究,为利用该模型进行其它研究提供实验依据。方法：选用2、4、6、8、10、12月龄SAMP8,以同龄正常老化的SAMR1为对照,应用老化度评分和Morris水迷宫测试SAMP8的老化度和学习记忆能力的增龄性变化。结果：SAMP8在4月龄出现明显的老化体征,并在6月龄开始表现出空间学习记忆障碍。结论：SAMP8在成熟期（4-6月）过后发生了快速老化,证实SAMP8是研究衰老和老年性痴呆的理想模型。     

卡那霉素耳慢性中毒对豚鼠耳蜗毛细胞中Bcl-2表达的影响

目的:探讨卡那霉素耳慢性中毒对豚鼠耳蜗毛细胞中Bcl-2表达的影响。方法:取20只豚鼠随机分为2组,实验组连续14d肌肉注射硫酸卡那霉素,200mg/(kg.d),对照组连续14d等量肌肉注射生理盐水,停药14d处死动物后制作耳蜗标本,处死前检测其ABR的变化,免疫组化及原位杂交法测定Bcl-2的表达。结果:豚鼠卡那霉素耳慢性中毒后,ABR阈值较对照组明显上升,Bcl-2阳性表达减低,与对照组比较差异有显著性(P<0.05)。结论:卡那霉素耳慢性损害可能与抑制Bcl-2的表达有关。     

Caspase-3在老年豚鼠耳蜗螺旋神经节细胞的表达

目的检测caspase-3在老年豚鼠耳蜗的表达。方法实验分两组:实验组和对照组,实验组豚鼠年龄为33至35个月之间,对照组豚鼠年龄为2至3个月。用免疫组织化学方法检测caspase-3在两组豚鼠耳蜗的表达。结果Caspase-3在实验组耳蜗的表达呈阳性,阳性区域主要存在于耳蜗螺旋神经节细胞。在对照组耳蜗的表达呈阴性。结论Caspase-3在老年豚鼠耳蜗螺旋神经节细胞中呈阳性表达,提示caspase-3在豚鼠耳蜗老化过程中起重要作用。     

猫耳蜗电图中N_2波起源的分析

在35只猫进行了耳蜗电图、听觉脑干电反应及耳蜗核局部电位的同时描记,将普鲁卡因或海人酸微量注入耳蜗核内,观察电位的变化,以分析耳蜗电图中N_2 波的起源。实验结果表明:猫的 N_2 波来源于外周第一级神经元冲动的成分和耳蜗核电活动的成分。     

种子老化过程中膜的变化及其原因

综述种子老化过程中细胞膜结构和透性的变化,分析种子老化过程中引起膜变化的原因。指出目前对种子老化和膜关系的研究现状及存在的问题。     

心梗后心肌重构过程中AT1A，AT2受体表达的变化

为探讨AT1,AT2受体在心肌重构演变过程中的作用,本实验应用免疫组化,电镜技术和图像分析方法,观察了大鼠心梗后心肌重构过程中非醒,AT1,AT2受体表达的动态变化,结果显示,心梗术后3d,电镜显示非梗塞区心肌细胞肌原纤维横纹消失,线粒体肿胀,成纤维细胞增多,免疫组化显示AT1A受体在非梗塞区心肌组织表达明显升高（P＜0．001）,AT2受体表达无明显变化（P＞0．05）,心梗术后14天,可见心肌细胞肌原纤维模纹,心肌细胞间胶原纤维明显增多。同时AT1A受本在心肌的表达比心梗术后3天时减弱,但仍高于对照组（P＜0．05）,AT2受体表达明显增加（P＜0．001）,结果提示：心梗后非梗塞区心肌AT1A,AT2受体表达先后上调,可能参与介导心肌重构过程。     

MeCP2与神经发育性疾病

作为一种转录抑制因子,甲基化CpG结合蛋白2（MeCP2）含有结合甲基化DNA和转录抑制两个特征性的结构域,具有调节转录激活、调节染色体构象、参与RNA剪切等多种功能,在神经发育过程中起着重要的作用。近来的研究表明,MeCP2基因突变与Rett综合征、孤独症等多种神经发育性疾病相关,已成为研究基因型与人类神经发育性疾病关系的一个热点。就MeCP2在Rett综合征、孤独症及药物成瘾方面的进展作一综述。     

细胞外基质的增龄性变化及分子机制的研究进展

人口老龄化及其伴随的各种疾病已成为全球性健康问题,细胞外基质在老化过程中发生的变化及对机体产生的影响逐渐成为研究衰老的热点。在机体发育和衰老过程中,细胞外基质不仅可以为细胞提供结构支架、组织连接,调节实质细胞的形态、增殖、分化、代谢、迁移等生理活动,并且其本身组成成分、合成、代谢、重构等变化也会对机体各系统的功能产生深刻影响,具体表现为骨骼肌僵硬、左心室功能受损、神经突触传导抑制等。本文通过介绍机体在衰老过程中,运动、循环、神经等系统细胞外基质的变化及相关机制的最新研究进展,从非细胞角度探讨老化的机制,了解衰老的过程。     

MeCP2 interacts with chromosomal microRNAs in brain

Although methyl CpG binding domain protein-2 (MeCP2) is commonly understood to function as a silencing factor at methylated DNA sequences, recent studies also show that MeCP2 can bind unmethylated sequences and coordinate gene activation. MeCP2 displays broad binding patterns throughout the genome, with high expression levels similar to histone H1 in neurons. Despite its significant presence in the brain, only subtle gene expression changes occur in the absence of MeCP2. This may reflect a more complex regulatory mechanism of MeCP2 to complement chromatin binding. Using an RNA immunoprecipitation of native chromatin technique, we identify MeCP2 interacting microRNAs in mouse primary cortical neurons. In addition, comparison with mRNA sequencing data from Mecp2-null mice suggests that differentially expressed genes may indeed be targeted by MeCP2-interacting microRNAs. These findings highlight the MeCP2 interaction with microRNAs that may modulate its binding with chromatin and regulate gene expression.     

Regulation and function of stimulus-induced phosphorylation of MeCP2

DNA methylation-dependent epigenetic regulation plays important roles in the development and function of the mammalian nervous system. MeCP2 is a key player in recognizing methylated DNA and interpreting the epigenetic information encoded in different DNA methylation patterns. Mutations in the MECP2 gene cause Rett syndrome, a devastating neurological disease that shares many features with autism. One interesting aspect of MeCP2 function is that it can be phosphorylated in response to diverse stimuli. Insights into the regulation and function of MeCP2 phosphorylation will help improve our understanding of how MeCP2 integrates environmental stimuli in neuronal nuclei to generate adaptive responses and may eventually lead to treatments for patients.     

SIRT1-mediated deacetylation of MeCP2 contributes to BDNF expression

Methyl-CpG binding protein 2 (MeCP2) binds methylated cytosines at CpG sites on DNA and it is thought to function as a critical epigenetic regulator. Mutations in the MeCP2 gene have been associated to Rett syndrome, a human neurodevelopmental disorder. Here we show that MeCP2 is acetylated by p300 and that SIRT1 mediates its deacetylation. SIRT1, the mammalian homologue of Sir2 in yeast, is a nicotinamide-adenine dinucleotide (NAD⁺)-dependent histone deacetylase that belongs to the family of HDAC class III sirtuins. Importantly, SIRT1 has been shown to play a critical role in synaptic plasticity and memory formation. This study reveals a functional interplay between two critical epigenetic regulators, MeCP2 and SIRT1, which controls MeCP2 binding activity to the brain-derived neurotrophic factor (BDNF) promoter in a specific region of the brain.     

SIRT1-mediated deacetylation of MeCP2 contributes to BDNF expression

Methyl-CpG binding protein 2 (MeCP2) binds methylated cytosines at CpG sites on DNA and it is thought to function as a critical epigenetic regulator. Mutations in the MeCP2 gene have been associated to Rett syndrome, a human neurodevelopmental disorder. Here we show that MeCP2 is acetylated by p300 and that SIRT1 mediates its deacetylation. SIRT1, the mammalian homologue of Sir2 in yeast, is a nicotinamide-adenine dinucleotide (NAD⁺)-dependent histone deacetylase that belongs to the family of HDAC class III sirtuins. Importantly, SIRT1 has been shown to play a critical role in synaptic plasticity and memory formation. This study reveals a functional interplay between two critical epigenetic regulators, MeCP2 and SIRT1, which controls MeCP2 binding activity to the brain-derived neurotrophic factor (BDNF) promoter in a specific region of the brain.     

Autistic-like social deficits in hippocampal MeCP2 knockdown rat models are rescued by ketamine

Autism or autism spectrum disorder (ASD) is a behavioral syndrome characterized by persistent deficits in social interaction, and repetitive patterns of behavior, interests, or activities. The gene encoding Methyl-CpG binding protein 2 (MeCP2) is one of a few exceptional genes of established causal effect in ASD. Although genetically engineered mice studies may shed light on how MeCP2 loss affects synaptic activity patterns across the whole brain, such studies are not considered practical in ASD patients due to the overall level of impairment, and are technically challenging in mice. For the first time, we show that hippo-campal MeCP2 knockdown produces behavioral abnormalities associated with autism-like traits in rats, providing a new strategy to investigate the efficacy of therapeutics in ASD. Ketamine, an N-Methyl-D-aspartate (NMDA) blocker, has been proposed as a possible treatment for autism. Using the MeCP2 knockdown rats in conjunction with a rat model of valproic acid (VPA)-induced ASD, we examined gene expression and ASD behaviors upon ketamine treatment. We report that the core symptoms of autism in MeCP2 knockdown rats with social impairment recovered dramatically following a single treatment with ketamine.