Data for: Visualizing Reactive Oxygen Species in live mammals

Published: 18 August 2019| Version 3 | DOI: 10.17632/jm3w3gv2px.3
Contributors:
Jingke Guo,
Huiqin Wang,
Xi Cheng,
Lijing Ke,
Qiang Wang,
Hang Xu,
Suhai Qiu,
Shusen Zheng,
Tianbao Chen,
Xin Zhang,
Zicai Wang,
Shutao Liu,
Lina Qin,
Yanglei Cheng,
Shuyu Zheng,
Guanzhen Gao,
Xicui Sun,
Wei Ding,
Yuqiu Hu,
Xiaowei Yu,
Mingming Xu,
Penghan Huang,
Qiming Wu,
Chuntong Lin,
You Dai,
Zhe Li,
Dazheng Xu,
Qi Chen,
Pingfan Rao,
Yingjie Lin,
Jianzhi Lu,
Yi Guo,
Jianwu Zhou,
Sherry Ho,
Chris Shaw,
Han Chen,
Huaiyu Gu,
Jiaqi Wang

Description

Visualising ROS distribution in Cynomolgus monkeys After anesthesia, 25 mg/kg DCFH-DA (Sigma-aldrich, St. Louis, Missouri, USA) and 2.5 mg/kg of MitoSOX (Thermo Fisher Scientific, Eugene, Oregon, USA) were dissolved in 5 ml/kg dimethyl sulfoxide (Sinopharm Chemical Reagent, Shanghai, China), then injected in small saphenous vein of Cynomolgus monkeys in 30 minutes. 30 minutes after injection, the skin was removed off the main anterior body to observe ROS distribution in forehead, face, chest, abdomen and perineum, and then the skull, thoracic cavity and abdominal cavity were open for the observation of relevant organs and tissues. The exposed parts were minutely incised. Immunohistochemical and microscopic investigation of the fluorescent linea alba was conducted on a sample of 1×1×3 mm after incubation with DAPI for visualizing cell nucleus, Cy3 for collagen, then observed with a transmission electron microscope (TEM, FEI, Valley City, ND, USA) at a voltage of 80 KV. Structural properties of the concentrated ROS distribution in linea alba of SD rats To identify elements responsible for the fluorescent light emission, linea alba of SD rats were studied. After anesthesia, 1 ml of DCFH-DA solution (5 mg in 1 ml DMSO) was injected into the tail vein. 30 minutes later, the frontal abdominal wall was incised, and further cross sectioned for the examination by a microscope (Leica, Buffalo Grove, Illinois, USA). Electric properties of the concentrated ROS distribution in linea alba of SD rats 1 ml of DCFH-DA solution (0.05 mg in 1 ml DMSO) was smeared directly on the linea alba on the interior surface of an isolated frontal abdominal wall with and without the reconnecting linea alba by electric wire to the cutting points (xiphoid and symphysis pubis). A digital ammeter was used to measure the current between the xiphoid and symphysis pubis on the linea alba. Measurement of electric current in linea alba of mice The mice were anesthetized. The abdominal walls of mice were incised. A digital ammeter was used to measure the current between the xiphoid and symphysis pubis on the linea alba. The symphysis pubis was marked with a positive terminal. The linea alba current was measured with three groups of mice: cpYFP- group: 6 male C57BL/6 mice, weight 22.7±0.8 g. Connect the body to an hour later, 300 μl Adeionized water drops were dripped on liver. cpYFP+ group: 6 male C57BL/6-TgH(mito-cpYFP) mice, weight 21.2±1.3 g. Connect the body to an hour later, 300 μl deionized water drops were dripped on liver. cpYFP+H2O2 group: 6 male C57BL/6-TgH(mito-cpYFP) mice, weight 21.3±1.5 g. Connect the body to an hour later, 300 μl 10mM H2O2 drops were dripped on liver.

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