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Identificação molecular e análise do padrão de expressão tecidual dos genes relacionados às sirtuínas em zebrafish

机译:斑马鱼中sirtuins相关基因的分子鉴定和组织表达模式分析

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摘要

First identified in Saccharomyces cerevisie as silent information regulators (SIRs) sirtuins (SIRTs) are widely distributed in all phyla of life. These large and diverse class III histone deacetylases (HDACs) comprise a family of NAD+-dependent protein deacetylases that catalyze mono-ADP-ribosyl transfer and/or deacetylation, and are considered a highly conserved family of proteins. Zebrafish (Danio rerio) is a freshwater fish widely used as model organism in many research areas. Characteristics such as low costs of breeding and maintenance, easy water-soluble drugs administration and rapid adaptation to new environments, among others, made the zebrafish a suitable model organism for studying many human diseases; including aging-associated disorders. Despite its promising use as a vertebrate model of aging, little is known about the presence of sirtuin-related genes in zebrafish, which has been considered an important research target for its therapeutics potential. Therefore, the purpose of this study was to identify all sirtuin-related genes and to map their expression patterns in nine different tissues of adult zebrafish. Total RNA was extracted from tissues samples of male and female zebrafish. RT-PCR reactions were performed under optimized conditions for expression pattern analysis of the sirtuins-related genes, previously identified using NCBI-Blast searches. Results showed the existence of eight sirtuins-related genes, including two paralogs. Each gene presented a unique expression pattern, illustrating their wide tissue distribution. In order to test a modulatory effect on gene expression of the sirtuin members, we exposed fish to resveratrol and the results demonstrated an alteration of the expression levels of some sirtuins. In this sense, we suggest that new drugs potentially able to modulate sirtuins expression could be tested using the system here described, aiming, in the future, to develop more selective therapies for age-associated disorders
机译:首先在酿酒酵母中被鉴定为沉默信息调节剂(SIR)沉默调节蛋白(SIRT)广泛分布于生命的所有门系中。这些庞大而多样的III类组蛋白脱乙酰基酶(HDAC)包含NAD +依赖性蛋白脱乙酰基酶家族,可催化​​单ADP-核糖基转移和/或脱乙酰基化,被认为是高度保守的蛋白家族。斑马鱼(Danio rerio)是一种淡水鱼,在许多研究领域中被广泛用作模型生物。斑马鱼的特点是诸如低廉的繁殖和维护成本,易于施用的水溶性药物以及快速适应新环境等特征,使其成为研究许多人类疾病的合适模式生物。包括与衰老相关的疾病。尽管它有希望用作衰老的脊椎动物模型,但对于斑马鱼中沉默调节蛋白相关基因的存在知之甚少,斑马鱼已被认为是其治疗潜力的重要研究目标。因此,本研究的目的是鉴定所有与sirtuin相关的基因,并绘制其在成年斑马鱼的九种不同组织中的表达模式。从雄性和雌性斑马鱼的组织样品中提取总RNA。 RT-PCR反应是在优化条件下进行的,用于sirtuins相关基因的表达模式分析,先前已使用NCBI-Blast搜索进行了鉴定。结果显示存在八个sirtuins相关基因,包括两个旁系同源物。每个基因均表现出独特的表达方式,说明了其广泛的组织分布。为了测试对sirtuin成员的基因表达的调节作用,我们将鱼暴露于白藜芦醇,结果证明了某些sirtuin的表达水平发生了变化。从这个意义上讲,我们建议可以使用此处描述的系统测试可能能够调节沉默调节蛋白表达的新药,以期在将来针对与年龄相关的疾病开发更多的选择性疗法

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