Lau Gigi Y, Barts Nicholas, Hartley Richard C, Tobler Michael, Richards Jeffrey G, Murphy Michael P, Arndt Sabine
Department of Zoology, University of British Columbia, 6270 University Boulevard, Vancouver, BC V6T 1Z4, Canada
Division of Biology, Kansas State University, 116 Ackert Hall, Manhattan, KS 66506, USA.
Biol Open. 2019 May 9;8(5):bio041467. doi: 10.1242/bio.041467.
In this paper, we outline the use of a mitochondria-targeted ratiometric mass spectrometry probe, MitoA, to detect changes in mitochondrial hydrogen sulfide (HS) in (family Poeciliidae). MitoA is introduced via intraperitoneal injection into the animal and is taken up by mitochondria, where it reacts with HS to form the product MitoN. The MitoN/MitoA ratio can be used to assess relative changes in the amounts of mitochondrial HS produced over time. We describe the use of MitoA in the fish species to illustrate the steps for adopting the use of MitoA in a new organism, including extraction and purification of MitoA and MitoN from tissues followed by tandem mass spectrometry. In this proof-of-concept study we exposed HS tolerant to 59 µM free HS for 5 h, which resulted in increased MitoN/MitoA in brain and gills, but not in liver or muscle, demonstrating increased mitochondrial HS levels in select tissues following whole-animal HS exposure. This is the first time that accumulation of HS has been observed during whole-animal exposure to free HS using MitoA. This article has an associated First Person interview with the first author of the paper.
在本文中，我们概述了一种线粒体靶向的比率质谱探针MitoA的用途，用于检测食蚊鱼（花鳉科）中线粒体硫化氢（HS）的变化。MitoA通过腹腔注射引入动物体内，并被线粒体摄取，在那里它与HS反应形成产物MitoN。MitoN/MitoA比率可用于评估随时间产生的线粒体HS量的相对变化。我们描述了MitoA在该鱼类中的使用，以说明在新生物体中采用MitoA的步骤，包括从组织中提取和纯化MitoA和MitoN，然后进行串联质谱分析。在这项概念验证研究中，我们将耐HS的食蚊鱼暴露于59μM游离HS中5小时，这导致大脑和鳃中的MitoN/MitoA增加，但肝脏或肌肉中没有增加，表明在全动物HS暴露后，特定组织中的线粒体HS水平升高。这是首次使用MitoA在全动物暴露于游离HS期间观察到HS的积累。本文配有对该论文第一作者的第一人称访谈。
