Whole organism and tissue specific analysis of pexophagy in Drosophila
Whole organism and tissue specific analysis of pexophagy in Drosophila
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果蝇自噬的整体和组织特异性分析
DOI：
10.1101/2023.11.17.567516
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Barone F
中科院分区：
文献类型：
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作者：
Barone F
过氧化物酶体是参与动物关键代谢过程的重要细胞器，例如脂肪酸氧化、醚磷脂产生和活性氧解毒。我们已经生成了转基因黑腹果蝇模型，表达过氧化物酶体选择性自噬的荧光报告基因，这一过程称为 pexophagy。我们表明这些报告基因与过氧化物酶体标记物共定位，并且它们可以反映铁螯合诱导的自噬诱导和核心自噬蛋白 Atg5 耗尽的抑制。使用光片显微镜，我们已经能够全面了解整个生物体在不同发育阶段的自噬水平。 pexophagy 的组织特异性控制的例子是过氧化物酶体丰富但 pexophagy 最小的区域，在被上皮细胞包围的卵细胞簇中观察到，其中 pexophagy 更加明显。通过给果蝇喂食铁螯合剂去铁酮来增强自噬能力，这与过去使用哺乳动物细胞的结果一致。使用特定的驱动程序来可视化神经元中的自噬，并证明幼虫中枢神经系统中 Hsc70-5（分子伴侣 HSPA9/mortalin 的果蝇同源物）的特定消耗导致了自噬的显着升高。
Peroxisomes are essential organelles involved in critical metabolic processes in animals such as fatty acid oxidation, ether phospholipid production and reactive oxygen species detoxification. We have generated transgenicDrosophila melanogastermodels expressing fluorescent reporters for the selective autophagy of peroxisomes, a process known as pexophagy. We show that these reporters are colocalized with a peroxisomal marker and that they can reflect pexophagy induction by iron chelation and inhibition by depletion of the core autophagy protein Atg5. Using light sheet microscopy, we have been able to obtain a global overview of pexophagy levels across the entire organism at different stages of development. Tissue-specific control of pexophagy is exemplified by areas of peroxisome abundance but minimal pexophagy, observed in clusters of oenocytes surrounded by epithelial cells where pexophagy is much more evident. Enhancement of pexophagy was achieved by feeding flies with the iron chelator deferiprone, in line with past results using mammalian cells. Specific drivers were used to visualize pexophagy in neurons, and to demonstrate that specific depletion in the larval central nervous system of Hsc70-5, theDrosophilahomologue of the chaperone HSPA9/mortalin, led to a substantial elevation in pexophagy.