Department of Molecular Biotechnology and Microbiology, Institute of Biotechnology, Faculty of Science and Technology, University of Debrecen, Egyetem tér 1, 4032, Debrecen, Hungary.
Department of Biophysics and Cell Biology, Faculty of Medicine, University of Debrecen, Egyetem tér 1, 4032, Debrecen, Hungary.
Appl Microbiol Biotechnol. 2022 Jun;106(11):3895-3912. doi: 10.1007/s00253-022-11967-2. Epub 2022 May 23.
While phototoxicity can be a useful therapeutic modality not only for eliminating malignant cells but also in treating fungal infections, mycologists aiming to observe morphological changes or molecular events in fungi, especially when long observation periods or high light fluxes are warranted, encounter problems owed to altered regulatory pathways or even cell death caused by various photosensing mechanisms. Consequently, the ever expanding repertoire of visible fluorescent protein toolboxes and high-resolution microscopy methods designed to investigate fungi in vitro and in vivo need to comply with an additional requirement: to decrease the unwanted side effects of illumination. In addition to optimizing exposure, an obvious solution is red-shifted illumination, which, however, does not come without compromises. This review summarizes the interactions of fungi with light and the various molecular biology and technology approaches developed for exploring their functions on the molecular, cellular, and in vivo microscopic levels, and outlines the progress towards reducing phototoxicity through applying far-red and near-infrared light. KEY POINTS: • Fungal biological processes alter upon illumination, also under the microscope • Red shifted fluorescent protein toolboxes decrease interference by illumination • Innovations like two-photon, lightsheet, and near IR microscopy reduce phototoxicity.
虽然光毒性不仅可用于消除恶性细胞,还是一种有用的治疗模式,也可用于治疗真菌感染,但真菌学家在观察真菌的形态变化或分子事件时,特别是在需要长时间观察或高光通量的情况下,会遇到各种光感机制引起的调控途径改变甚至细胞死亡的问题。因此,不断扩展的可见荧光蛋白工具盒和高分辨率显微镜方法 repertoire 旨在研究体外和体内真菌,需要符合额外的要求:减少不必要的光照副作用。除了优化曝光之外,一个明显的解决方案是红移照射,但这并非没有妥协。本综述总结了真菌与光的相互作用,以及为探索其在分子、细胞和活体微观水平上的功能而开发的各种分子生物学和技术方法,并概述了通过应用远红光和近红外光来降低光毒性的进展。要点:
真菌的生物学过程在光照下发生变化,即使在显微镜下也是如此;
红移荧光蛋白工具盒减少了光照的干扰;
双光子、光片和近红外显微镜等创新技术降低了光毒性。