The defective expression of gtpbp3 related to tRNA modification alters the mitochondrial function and development of zebrafish
The defective expression of gtpbp3 related to tRNA modification alters the mitochondrial function and development of zebrafish
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与 tRNA 修饰相关的 gtpbp3 表达缺陷改变了斑马鱼的线粒体功能和发育
DOI：
10.1016/j.biocel.2016.05.012
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发表时间：
2016-08-01
影响因子：
4
通讯作者：
Guan, Min-Xin
中科院分区：
文献类型：
--
作者：
Chen, Danni;Li, Feng;Guan, Min-Xin
人类线粒体 DNA (mtDNA) 突变与多种临床异常相关。然而，核修饰基因调节致病性 mtDNA 突变的表型表达。在我们之前的研究中，我们发现了与线粒体 tRNA 修饰相关的人类 GTPBP3，作为影响耳聋相关 mtDNA 突变的修饰剂。 GTPBP3 突变已被发现与其他人类疾病有关。然而，GTPBP3 相关疾病的病理生理学仍未完全了解。在这里，我们报道了使用反义吗啉代的 Gtpbp3 耗竭斑马鱼模型的生成和表征。斑马鱼 gtpbp3 具有位于线粒体的三种亚型。斑马鱼 gtpbp3 在不同胚胎阶段和多种组织中表达。特别是，gtpbp3 在成年斑马鱼卵巢和睾丸中表达更丰富。斑马鱼gtpbp3的表达可以功能性地恢复酵母中mss1/gtpbp3突变引起的生长缺陷。在 gtpbp3 敲低的斑马鱼胚胎中观察到线粒体 ATP 生成显着减少，同时细胞凋亡和活性氧水平增加。与未注射的对照相比，Gtpbp3 突变体在受精后 5 天内表现出胚胎发育缺陷，包括出血、黑色素、水肿和尾巴弯曲。野生型 gtpbp3 mRNA 的共注射部分挽救了 Gtpbp3 突变体中的这些缺陷。这些数据表明斑马鱼 Gtpbp3 是人类和酵母 GTPBP3 的结构和功能同源物。 Gtpbp3 缺陷引起的线粒体功能障碍可能会改变斑马鱼的胚胎发育。此外，这种线粒体疾病斑马鱼模型可能为研究缺陷 tRNA 修饰、线粒体生物发生和线粒体疾病的病理生理学提供独特的机会。 (C) 2016 Elsevier Ltd. 保留所有权利。
Human mitochondrial DNA (mtDNA) mutations have been associated with a wide spectrum of clinical abnormalities. However, nuclear modifier gene(s) modulate the phenotypic expression of pathogenic mtDNA mutations. In our previous investigation, we identified the human GTPBP3 related to mitochondrial tRNA modification, acting as a modifier to influence of deafness-associated mtDNA mutation. Mutations in GTPBP3 have been found to be associated with other human diseases. However, the pathophysiology of GTPBP3-associated disorders is still not fully understood. Here, we reported the generation and characterization of Gtpbp3 depletion zebrafish model using antisense morpholinos. Zebrafish gtpbp3 has three isoforms localized at mitochondria. Zebrafish gtpbp3 is expressed at various embryonic stages and in multiple tissues. In particular, the gtpbp3 was expressed more abundantly in adult zebrafish ovary and testis. The expression of zebrafish gtpbp3 can functionally restore the growth defects caused by the mss1/gtpbp3 mutation in yeast. A marked decrease of mitochondrial ATP generation accompanied by increased levels of apoptosis and reactive oxygen species were observed in gtpbp3 knockdown zebrafish embryos. The Gtpbp3 morphants exhibited defective in embryonic development including bleeding, melenin, oedema and curved tails within 5 days post fertilization, as compared with uninjected controls. The co-injection of wild type gtpbp3 mRNA partially rescued these defects in Gtpbp3 morphants. These data suggest that zebrafish Gtpbp3 is a structural and functional homolog of human and yeast GTPBP3. The mitochondrial dysfunction caused by defective Gtpbp3 may alter the embryonic development in the zebrafish. In addition, this zebrafish model of mitochondrial disease may provide unique opportunities for studying defective tRNA modification, mitochondrial biogenesis, and pathophysiology of mitochondrial disorders. (C) 2016 Elsevier Ltd. All rights reserved.