Visualizing Lipid Droplets in Drosophila Oogenesis.
Visualizing Lipid Droplets in Drosophila Oogenesis.
复制标题
果蝇卵子发生过程中脂滴的可视化。
DOI：
10.1007/978-1-0716-2970-3_12
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Welte,MichaelA
中科院分区：
文献类型：
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作者：
White,RogerP;Welte,MichaelA
脂滴是一种在脊椎动物和无脊椎动物卵母细胞和卵子中含量丰富的脂肪储存细胞器。它们在卵子发生和提供发育中的胚胎中都有作用。在果蝇中，大量的LD在卵子发生中期的营养细胞中产生，然后转移到卵母细胞中。它们的数量和空间分布随着发育和对各种实验操作的反应而变化。本章演示了如何在固定组织和活样本中观察果蝇滤泡中的LD。对于固定样本，方案解释了如何准备雌蝇、解剖卵巢、分离卵泡、固定、应用染色剂、固定组织和进行成像。对于活体样本，该协议显示了如何解剖卵巢，应用荧光LD染料，培养卵泡，使它们在成像过程中保持活力和健康。最后，提供了一种采用体内离心来评估标记物与LD的共定位的方法。
Lipid droplets (LDs) are fat storage organelles highly abundant in oocytes and eggs of many vertebrates and invertebrates. They have roles both during oogenesis and in provisioning the developing embryo. InDrosophila, large numbers of LDs are generated in nurse cells during mid-oogenesis and then transferred to oocytes. Their number and spatial distribution changes developmentally and in response to various experimental manipulations. This chapter demonstrates how to visualize LDs inDrosophilafollicles, both in fixed tissues and living samples. For fixed samples, the protocol explains how to prepare female flies, dissect ovaries, isolate follicles, fix, apply stains, mount the tissue, and perform imaging. For live samples, the protocol shows how to dissect ovaries, apply a fluorescent LD dye, and culture follicles such that they remain alive and healthy during imaging. Finally, a method is provided that employs in vivo centrifugation to assess colocalization of markers with LDs.