Tools Allowing Independent Visualization and Genetic Manipulation of Drosophila melanogaster Macrophages and Surrounding Tissues.
Tools Allowing Independent Visualization and Genetic Manipulation of Drosophila melanogaster Macrophages and Surrounding Tissues.
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DOI：
10.1534/g3.117.300452
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发表时间：
2018-03-02
期刊：
影响因子：
--
通讯作者：
Siekhaus DE
中科院分区：
文献类型：
--
作者：
Gyoergy A;Roblek M;Ratheesh A;Valoskova K;Belyaeva V;Wachner S;Matsubayashi Y;Sánchez-Sánchez BJ;Stramer B;Siekhaus DE
果蝇黑腹浆细胞是血细胞中的吞噬细胞，对免疫反应至关重要，但也通过与其他类型细胞的相互作用发挥着从早期发育到死亡的关键作用。它们在发育、干细胞增殖、新陈代谢、癌症、创伤反应和衰老过程中调节动态平衡和信号传递，显示出与脊椎动物巨噬细胞有趣的分子和功能保守。考虑到与脊椎动物相比，果蝇的遗传学相对容易，允许在所有阶段独立地对浆细胞和周围组织进行可视化和遗传操作的工具将极大地帮助更全面地了解这些过程，但还缺乏。在这里，我们描述了一套全面的转基因品系来实现这一点。其中包括基于mCherry的极其明亮的荧光品系，它允许在活的和固定的样本中，甚至作为杂合子，对从胚胎阶段8到成年期的浆细胞核、细胞质或肌动蛋白细胞骨架进行GAL4独立的可视化，从而极大地促进了筛查。这些线允许实时可视化和跟踪胚胎浆细胞，以及驻留在体壁或与周围血淋巴一起流动的幼虫浆细胞。通过共聚焦成像，可以在活的或固定的胚胎、幼虫和成体中看到浆细胞和内部组织的相互作用。它们允许在整个生命周期内对浆细胞进行高效的GAL4非依赖荧光激活细胞分类(FACS)分析/分选。为了便于相互信号的遗传学研究，我们还建立了一个表达浆细胞的QF2系，与现有的GAL4驱动程序相结合，允许对浆细胞和周围组织进行独立的遗传操作，以及GAL80株，它阻止GAL4驱动程序影响浆细胞，所有这些都从早期胚胎发挥作用到成年。
Drosophila melanogaster plasmatocytes, the phagocytic cells among hemocytes, are essential for immune responses, but also play key roles from early development to death through their interactions with other cell types. They regulate homeostasis and signaling during development, stem cell proliferation, metabolism, cancer, wound responses, and aging, displaying intriguing molecular and functional conservation with vertebrate macrophages. Given the relative ease of genetics in Drosophila compared to vertebrates, tools permitting visualization and genetic manipulation of plasmatocytes and surrounding tissues independently at all stages would greatly aid a fuller understanding of these processes, but are lacking. Here, we describe a comprehensive set of transgenic lines that allow this. These include extremely brightly fluorescing mCherry-based lines that allow GAL4-independent visualization of plasmatocyte nuclei, the cytoplasm, or the actin cytoskeleton from embryonic stage 8 through adulthood in both live and fixed samples even as heterozygotes, greatly facilitating screening. These lines allow live visualization and tracking of embryonic plasmatocytes, as well as larval plasmatocytes residing at the body wall or flowing with the surrounding hemolymph. With confocal imaging, interactions of plasmatocytes and inner tissues can be seen in live or fixed embryos, larvae, and adults. They permit efficient GAL4-independent Fluorescence-Activated Cell Sorting (FACS) analysis/sorting of plasmatocytes throughout life. To facilitate genetic studies of reciprocal signaling, we have also made a plasmatocyte-expressing QF2 line that, in combination with extant GAL4 drivers, allows independent genetic manipulation of both plasmatocytes and surrounding tissues, and GAL80 lines that block GAL4 drivers from affecting plasmatocytes, all of which function from the early embryo to the adult.
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影响因子：
9.8
作者：
Crozatier M;Ubeda JM;Vincent A;Meister M
通讯作者：
Meister M
影响因子：
4.5
作者：
Chakrabarti S;Dudzic JP;Li X;Collas EJ;Boquete JP;Lemaitre B
通讯作者：
Lemaitre B
影响因子：
9.2
作者：
Clark, Rebecca I.;Woodcock, Katie J.;Geissmann, Frederic;Trouillet, Celine;Dionne, Marc S.
通讯作者：
Dionne, Marc S.
果蝇先天免疫：酚氧化酶原的区域和功能专业化。
影响因子：
5.4
作者：
Dudzic JP;Kondo S;Ueda R;Bergman CM;Lemaitre B
通讯作者：
Lemaitre B
DOI：
10.1084/jem.20060370
发表时间：
2006-11-27
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Greenberg ME;Sun M;Zhang R;Febbraio M;Silverstein R;Hazen SL
通讯作者：
Hazen SL