Visualizing Phagocytic Cargo In Vivo from Engulfment to Resolution in Caenorhabditis elegans.
Visualizing Phagocytic Cargo In Vivo from Engulfment to Resolution in Caenorhabditis elegans.
复制标题
可视化秀丽隐杆线虫体内吞噬细胞从吞噬到溶解的过程。
DOI：
10.1007/978-1-0716-3338-0_22
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Wehman,AnnM
中科院分区：
文献类型：
--
作者：
Fazeli,Gholamreza;Frondoni,Julia;Kolli,Shruti;Wehman,AnnM
线虫秀丽线虫在研究吞噬和清除吞噬的保守机制方面具有许多实验优势。这些问题包括用于时间推移成像的体内吞噬事件的刻板时间，标记参与吞噬作用不同步骤的分子的转基因记者的可用性，以及用于荧光成像的动物的透明度。此外，正向和反向遗传学公司的易用性。线虫使许多与吞噬清除有关的蛋白质的最初发现成为可能。在这一章中，我们关注的是大的未分化的卵裂球的吞噬作用。优雅胚胎，吞噬并清除从第二极体身体到细胞动学中体残留物的各种吞噬货物。我们描述了使用荧光延时成像来观察吞噬细胞清除的不同步骤，以及将这一过程正常化以区分突变菌株中的缺陷的方法。这些方法使我们能够从最初诱导吞噬的信号中揭示新的见解，直到吞噬溶酶体中的吞噬货物最终分解。
The nematodeCaenorhabditis elegansoffers many experimental advantages to study conserved mechanisms of phagocytosis and phagocytic clearance. These include the stereotyped timing of phagocytic events in vivo for time-lapse imaging, the availability of transgenic reporters labeling molecules involved in different steps of phagocytosis, and the transparency of the animal for fluorescence imaging. Further, the ease of forward and reverse genetics inC. eleganshas enabled many of the initial discoveries of proteins involved in phagocytic clearance. In this chapter, we focus on phagocytosis by the large undifferentiated blastomeres ofC. elegansembryos, which engulf and eliminate diverse phagocytic cargo from the corpse of the second polar body to cytokinetic midbody remnants. We describe the use of fluorescent time-lapse imaging to observe the distinct steps of phagocytic clearance and methods to normalize this process to distinguish defects in mutant strains. These approaches have enabled us to reveal new insights from the initial signaling to induce phagocytosis up until the final resolution of phagocytic cargo in phagolysosomes.