Balachandra Shruthi, Amodeo Amanda A
Department of Biological Sciences, Dartmouth College, Hanover, NH 03755, USA.
G3 (Bethesda). 2025 Jan 8;15(1). doi: 10.1093/g3journal/jkae271.
Quantitative live imaging is a valuable tool that offers insights into cellular dynamics. However, many fundamental biological processes are incompatible with current live-imaging modalities. Drosophila oogenesis is a well-studied system that has provided molecular insights into a range of cellular and developmental processes. The length of the oogenesis, coupled with the requirement for inputs from multiple tissues, has made long-term culture challenging. Here, we have developed Bellymount-pulsed tracking (Bellymount-PT), which allows continuous, noninvasive live imaging of Drosophila oogenesis inside the female abdomen for up to 16 h. Bellymount-PT improves upon the existing Bellymount technique by adding pulsed anesthesia with periods of feeding that support the long-term survival of flies during imaging. Using Bellymount-PT, we measure key events of oogenesis, including egg chamber growth, yolk uptake, and transfer of specific proteins to the oocyte during nurse cell dumping with high spatiotemporal precision within the abdomen of a live female.
定量活体成像技术是一种有价值的工具，可用于深入了解细胞动态。然而，许多基本生物学过程与当前的活体成像方式不兼容。果蝇卵子发生是一个经过充分研究的系统，为一系列细胞和发育过程提供了分子层面的见解。卵子发生的时长，再加上需要多个组织的输入，使得长期培养具有挑战性。在此，我们开发了腹部固定脉冲追踪技术（Bellymount-PT），该技术可对雌性果蝇腹部内的卵子发生过程进行长达16小时的连续、非侵入性活体成像。Bellymount-PT在现有的腹部固定技术基础上进行了改进，增加了脉冲麻醉和喂食周期，以支持果蝇在成像过程中的长期存活。使用Bellymount-PT，我们能够在活体雌性果蝇的腹部内，以高时空精度测量卵子发生的关键事件，包括卵室生长、卵黄摄取以及在营养细胞倾卸期间特定蛋白质向卵母细胞的转移。
