DNA damage signaling in Drosophila macrophages modulates systemic cytokine levels in response to oxidative stress

Environmental factors, infection, or injury can cause oxidative stress in diverse tissues and loss of tissue homeostasis. Effective stress response cascades, conserved from invertebrates to mammals, ensure reestablishment of homeostasis and tissue repair. Hemocytes, the Drosophila blood-like cells, rapidly respond to oxidative stress by immune activation. However, the precise signals how they sense oxidative stress and integrate these signals to modulate and balance the response to oxidative stress in the adult fly are ill-defined. Furthermore, hemocyte diversification was not explored yet on oxidative stress. Here, we employed high throughput single nuclei RNA-sequencing to explore hemocytes and other cell types, such as fat body, during oxidative stress in the adult fly. We identified distinct cellular responder states in plasmatocytes, the Drosophila macrophages, associated with immune response and metabolic activation upon oxidative stress. We further define oxidative stress-induced..., Drosophila melanogaster stocks: Flies were reared on a high yeast food containing 10% brewer’s yeast, 8% fructose, 2% polenta and 0.8% Agar. Propionic acid and nipagin were added to prevent bacterial or fungal growth. All crosses (except those containing the tub-Gal80ts construct or otherwise noted) were performed at 25°C with a 12 hours dark/light cycle. The crosses with tub-Gal80ts were performed at 18°C to ensure the inhibition of the Gal4-protein during developmental stages of the fly. The experimental male F1 flies were transferred to 29°C as soon as they hatched and were aged for six days. All transgenic lines used in this study are listed below: List of Drosophila lines used in this study Source w1118;Hml∆ -Gal4,UAS-2xeGFP Gift from Marc S. Dionne w1118;UAS-tefu-IR Vienna Drosophila Research Center w1118;UAS-mei41-IR Vienna Drosophila Research Center w1118;UAS-nbs-IR Vienna Drosophila Research Center w1118;UAS-mei41-IR;UAS-tefu-IR made for this study w1118;UAS-..., , # Drosophila macrophage response to oxidative stress The data deposited here includes survival data, imaging analysis, glucose/glycogen measurements, qPCR assays as well as TLC to measure triglyerides. The precise methods and analyses are described in the method section of this dataset. ## Description of the data and file structure The data files deposited here include the following datasets: Fly survivals were performed as indicated in the method section. Genotypes and treatment are included in the raw counting files below. Dead flies are indicated as 1 in the table living flies are indicated as 0. *Survival data (Lifespan, Paraquat, Starvation)* Paraquat survival: Fig.1_S1A_PQ survival_2mM_15mM_30mM_raw data.xml: Fig.1I_crq-reaper 15mM PQ survival raw data.txt Fig.5_S1E_Hml;tubGal80_DDR-IR_survival15mMPQ_raw data.xml Fig.5A_15mM PQ survival Hml-Gal4_DDR-IR.txt Fig.6A_Hml-upd3 and Hml-upd3-IR 15mM PQ survival_raw data.xml Fig.6C_Hml-upd3 and Hml-upd3-IR 15mM PQ hemocyte q...环境因素、感染或损伤可诱发多种组织产生氧化应激，并导致组织稳态失衡。从无脊椎动物到哺乳动物均保守存在的有效应激应答通路，可确保机体重建稳态并完成组织修复。 果蝇的类血细胞（hemocytes）可通过免疫激活快速响应氧化应激。然而，在成年果蝇中，这类细胞感知氧化应激的精确信号通路，以及整合信号以调控、平衡氧化应激应答的机制仍未明确。此外，学界尚未探究氧化应激对果蝇类血细胞分化的影响。 本研究采用高通量单细胞核RNA测序（single nuclei RNA-sequencing）技术，探究成年果蝇在氧化应激状态下的类血细胞及脂肪体等其他细胞类型。我们在果蝇巨噬细胞——浆细胞（plasmatocytes）——中鉴定出与氧化应激下免疫应答及代谢激活相关的不同细胞应答状态。本研究进一步明确了氧化应激诱导的……（原文此处截断）。 黑腹果蝇（Drosophila melanogaster）品系相关信息： 实验果蝇饲养于高酵母培养基中，该培养基配方为：10%啤酒酵母、8%果糖、2%玉米粉及0.8%琼脂。添加丙酸与尼泊金酯以抑制细菌及真菌污染。所有杂交实验（除携带tub-Gal80ts元件或另有说明的组合外）均在25℃、12小时明暗循环条件下完成。携带tub-Gal80ts的杂交实验则在18℃下进行，以确保在果蝇发育阶段抑制Gal4蛋白的活性。实验用雄性F1代果蝇羽化后即刻转移至29℃环境，培养6天。本研究使用的所有转基因果蝇品系如下： 本研究使用的果蝇品系列表及来源： w1118;Hml∆-Gal4,UAS-2xeGFP：由Marc S. Dionne教授惠赠 w1118;UAS-tefu-IR：维也纳果蝇研究中心 w1118;UAS-mei41-IR：维也纳果蝇研究中心 w1118;UAS-nbs-IR：维也纳果蝇研究中心 w1118;UAS-mei41-IR;UAS-tefu-IR：本研究定制构建 w1118;UAS-……，，# 果蝇巨噬细胞对氧化应激的应答 本数据集上传的数据涵盖生存实验数据、成像分析结果、葡萄糖/糖原含量检测数据、实时荧光定量PCR（qPCR）实验结果，以及用于测定甘油三酯含量的薄层色谱（TLC）数据。具体实验方法与数据分析流程详见本数据集的方法学部分。 数据与文件结构说明： 本数据集上传的数据文件包含以下数据集： 果蝇生存实验按照方法学部分所述流程开展，基因型与处理条件详见下方原始计数文件。表格中死亡果蝇标记为1，存活果蝇标记为0。 * 生存数据（寿命、百草枯处理、饥饿处理） 百草枯生存实验： Fig.1_S1A_PQ survival_2mM_15mM_30mM_raw data.xml Fig.1I_crq-reaper 15mM PQ survival raw data.txt Fig.5_S1E_Hml;tubGal80_DDR-IR_survival15mMPQ_raw data.xml Fig.5A_15mM PQ survival Hml-Gal4_DDR-IR.txt Fig.6A_Hml-upd3 and Hml-upd3-IR 15mM PQ survival_raw data.xml Fig.6C_Hml-upd3 and Hml-upd3-IR 15mM PQ hemocyte q……（原文此处截断）