Time-Lapse Imaging of Necroptosis and DAMP Release at Single-Cell Resolution
Time-Lapse Imaging of Necroptosis and DAMP Release at Single-Cell Resolution
复制标题
单细胞分辨率的坏死性凋亡和 DAMP 释放的延时成像
DOI：
10.1007/978-1-0716-1258-3_29
复制
发表时间：
2021
影响因子：
--
通讯作者：
Nakano Hiroyasu
中科院分区：
文献类型：
--
作者：
Murai Shin;Shirasaki Yoshitaka;Nakano Hiroyasu
坏死性上睑下垂是一种受调控的坏死形式，依赖于受体相互作用蛋白激酶（RIPK）3和混合谱系激酶结构域样蛋白（MLKL）。坏死细胞释放多种细胞和核因子，称为危险相关分子模式（DAMPs）。我们最近开发了一种förster共振能量转移（FRET）生物传感器，称为SMART（基于FRET的MLKL激活传感器）。SMART包含MLKL片段，它监测坏死性下垂，但不监测细胞凋亡或坏死。我们采用活细胞成像检测分泌活性（LCI-S），以单细胞分辨率观察坏死细胞高迁移率组盒1 （HMGB1）的释放。此外，我们结合SMART和lc - s成像技术，发现了两种不同的HMGB1在坏死细胞中的释放模式。因此，SMART和LCI-S是在单细胞分辨率下研究坏死性下垂和DAMP释放之间的密切相互作用的有价值的工具。
Necroptosis is a regulated form of necrosis that depends on receptor-interacting protein kinase (RIPK)3 and mixed lineage kinase domain-like protein (MLKL). Necroptotic cells release a variety of cellular and nuclear factors, referred to as danger-associated molecular patterns (DAMPs). We recently developed a förster resonance energy transfer (FRET) biosensor, termed SMART (a sensor for MLKL activation based on FRET). SMART comprises a fragment of MLKL, and it monitors necroptosis, but not apoptosis or necrosis. We performed live-cell imaging for secretion activity (LCI-S) to observe the release of high-mobility group box 1 (HMGB1) from necroptotic cells at single-cell resolution. Moreover, we combined SMART and LCI-S imaging techniques and found two different modes of HMGB1 release from necroptotic cells. Thus, SMART and LCI-S are valuable tools for investigating intimate cross talk between necroptosis and DAMP release at single-cell resolution.