Visualizing the Effects of Oxidative Damage on Drosophila Egg Chambers using Live Imaging.
Visualizing the Effects of Oxidative Damage on Drosophila Egg Chambers using Live Imaging.
复制标题
DOI：
10.3791/62157
复制
发表时间：
2021-04-10
期刊：
影响因子：
--
通讯作者：
Cox RT
中科院分区：
文献类型：
--
作者：
Sheard KM;Cox RT
果蝇卵巢的实时成像有助于了解发育过程中的各种基本细胞过程，包括核糖核蛋白颗粒运动，mRNA定位，细胞器运动和细胞骨架动力学。已经开发了几种用于实时成像的方法。由于每种方法都涉及解剖置于培养基或卤化碳油中的单个卵巢管，因此随着时间的推移，将不可避免地发生由于缺氧和/或物理操作引起的细胞损伤。缺氧的一个下游效应是增加细胞中的氧化损伤。本协议的目的是使用实时成像可视化的影响，在果蝇卵巢的亚细胞结构的定位和动力学诱导后，控制细胞损伤的氧化损伤。在这里，我们使用过氧化氢诱导细胞的氧化损伤，并给出这样的损伤对两个亚细胞结构，线粒体和Clu布利斯粒子的影响的例子。然而，这种方法适用于任何亚细胞结构。其局限性在于，过氧化氢只能添加到水性介质中，并且不能用于使用卤化碳油的成像。优点是过氧化氢容易获得且便宜，作用迅速，其浓度可以调节，并且氧化损伤是由缺氧引起的损伤以及由于操作引起的一般组织损伤的良好近似。本协议的目的是使用实时成像可视化的影响，在果蝇卵巢的亚细胞结构的定位和动态的氧化损伤。
Live imaging of Drosophila melanogaster ovaries has been instrumental in understanding a variety of basic cellular processes during development, including ribonucleoprotein particle movement, mRNA localization, organelle movement, and cytoskeletal dynamics. There are several methods for live imaging that have been developed. Due to the fact that each method involves dissecting individual ovarioles placed in media or halocarbon oil, cellular damage due to hypoxia and/or physical manipulation will inevitably occur over time. One downstream effect of hypoxia is to increase oxidative damage in the cells. The purpose of this protocol is to use live imaging to visualize the effects of oxidative damage on the localization and dynamics of subcellular structures in Drosophila ovaries after induction of controlled cellular damage. Here, we use hydrogen peroxide to induce cellular oxidative damage and give examples of the effects of such damage on two subcellular structures, mitochondria and Clu bliss particles. However, this method is applicable to any subcellular structure. The limitations are that hydrogen peroxide can only be added to aqueous media and would not work for imaging that uses halocarbon oil. The advantages are that hydrogen peroxide is readily available and inexpensive, acts quickly, its concentrations can be modulated, and oxidative damage is a good approximation of damage caused by hypoxia as well as general tissue damage due to manipulation. The objective of this protocol is to use live imaging to visualize the effects of oxidative damage on the localization and dynamics of subcellular structures in Drosophila ovaries.
登录
查看更多内容
影响因子：
64.8
作者：
WANG, SX;HAZELRIGG, T
通讯作者：
HAZELRIGG, T
影响因子：
4.3
作者：
Cox, Rachel T.;Spradling, Allan C.
通讯作者：
Spradling, Allan C.
影响因子：
2.4
作者：
Sen, Aditya;Cox, Rachel T.
通讯作者：
Cox, Rachel T.
影响因子：
2.7
作者：
Hudson, Andrew M.;Petrella, Lisa N.;Cooley, Lynn
通讯作者：
Cooley, Lynn
影响因子：
3.5
作者：
Winterbourn, CC
通讯作者：
Winterbourn, CC