Center for Clinical, Experimental Surgery, & Translational Research, Biomedical Research Foundation, Academy of Athens, 11527 Athens, Greece.
Laboratory of Pharmacology, Faculty of Pharmacy, National and Kapodistrian University of Athens, 15771 Athens, Greece.
Biomolecules. 2020 Feb 17;10(2):317. doi: 10.3390/biom10020317.
3-mercaptopyruvate sulfurtransferase (3-MST) is an enzyme capable of synthesizing hydrogen sulfide (HS) and polysulfides. In spite of its ubiquitous presence in mammalian cells, very few studies have investigated its contribution to homeostasis and disease development, thus the role of 3-MST remains largely unexplored. Here, we present a clustered, regularly interspaced, short palindromic repeats (CRISPR)/CRISPR-associated protein-9 (Cas9) induced mutant zebrafish line, which will allow the study of 3-MST's role in several biological processes. The zebrafish orthologue was identified using a bioinformatic approach and verified by its ability to produce HS in the presence of 3-mercaptopyruvate (3-MP). Its expression pattern was analyzed during zebrafish early development, indicating predominantly an expression in the heart and central nervous system. As expected, no detectable levels of 3-Mst protein were observed in homozygous mutant larvae. In line with this, HS levels were reduced in zebrafish. Although the mutants showed no obvious morphological deficiencies, they exhibited increased lethality under oxidative stress conditions. The elevated levels of reactive oxygen species, detected following deletion, are likely to drive this phenotype. In line with the increased ROS, we observed accelerated fin regenerative capacity in deficient zebrafish. Overall, we provide evidence for the expression of in zebrafish, confirm its important role in redox homeostasis and indicate the enzyme's possible involvement in the regeneration processes.
3-巯基丙酮酸硫转移酶（3-MST）是一种能够合成硫化氢（HS）和多硫化物的酶。尽管它广泛存在于哺乳动物细胞中，但很少有研究调查其对体内平衡和疾病发展的贡献，因此 3-MST 的作用在很大程度上仍未得到探索。在这里，我们展示了一个聚集的、规则间隔的、短的回文重复序列（CRISPR）/CRISPR 相关蛋白-9（Cas9）诱导的突变斑马鱼品系，这将允许研究 3-MST 在几个生物学过程中的作用。通过生物信息学方法鉴定了斑马鱼的同源物，并通过其在 3-巯基丙酮酸（3-MP）存在下产生 HS 的能力来验证。分析了其在斑马鱼早期发育过程中的表达模式，表明主要在心脏和中枢神经系统表达。不出所料，在纯合突变幼虫中未检测到 3-Mst 蛋白的可检测水平。与此一致，HS 水平在 斑马鱼中降低。尽管突变体没有明显的形态缺陷，但它们在氧化应激条件下的致死率增加。删除后检测到的活性氧水平升高可能导致这种表型。与增加的 ROS 一致，我们观察到 缺乏的斑马鱼中鳍再生能力的加速。总的来说，我们提供了 在斑马鱼中表达的证据，证实了它在氧化还原平衡中的重要作用，并表明该酶可能参与再生过程。
