Ceramide-rich microdomains facilitate nuclear envelope budding for non-conventional exosome formation.
Ceramide-rich microdomains facilitate nuclear envelope budding for non-conventional exosome formation.
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富含神经酰胺的微结构域促进非常规外泌体形成的核膜出芽。
DOI：
10.1038/s41556-022-00934-8
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发表时间：
2022-07
影响因子：
21.3
通讯作者：
中科院分区：
文献类型：
--
作者：
中性粒细胞向趋化梯度迁移，通过释放次级趋化因子白三烯B4(LTB4)来扩大其募集范围。我们先前已经证明LTB4及其合成酶5-脂氧合酶(5-LO)、5-LO激活蛋白(FLAP)和白三烯A4水解酶(LTA4H)被包装在外体中并被释放。我们现在报告，含有LTB4的外切体的生物发生起源于激活的中性粒细胞的核膜(NE)。我们发现中性鞘磷脂酶1(NSMase1)介导的神经酰胺富集脂有序微域的产生启动了LTB4合成酶在NE上的聚集。我们从激活的中性粒细胞中分离和分析了外切体，发现Flat/5-LO阳性的外切体群体与CD63阳性的外切体群体是不同的。此外，我们观察到Alix和Flat在细胞核外围和胞浆小泡内有很强的共存。我们认为，NE曲率和芽的形成是由nSMase1依赖的神经酰胺产生介导的，nSMase1依赖神经酰胺的产生导致瓣和Alix的募集。综上所述，这些观察阐明了LTB4分泌的机制，并确定了一条非传统的外切体产生途径。
Neutrophils migrating towards chemoattractant gradients amplify their recruitment range by releasing the secondary chemoattractant leukotriene B4 (LTB4). We previously demonstrated that LTB4 and its synthesizing enzymes, the 5-lipoxygenase (5-LO), 5-LO activating protein (FLAP), and leukotriene A4 hydrolase (LTA4H), are packaged and released in exosomes. We now report that the biogenesis of the LTB4-containing exosomes originates at the nuclear envelope (NE) of activated neutrophils. We show that the neutral sphingomyelinase 1 (nSMase1)-mediated generation of ceramide enriched lipid-ordered microdomains initiates the clustering of the LTB4-synthesizing enzymes on the NE. We isolated and analyzed exosomes from activated neutrophils and established that the FLAP/5-LO-positive exosome population is distinct from that of the CD63-positive exosome population. Furthermore, we observed a strong co-localization between ALIX and FLAP at the periphery of nuclei and within cytosolic vesicles. We propose that the initiation of NE curvature and bud formation is mediated by nSMase1-dependent ceramide generation, which leads to FLAP and ALIX recruitment. Together, these observations elucidate the mechanism for LTB4 secretion and identify a non-conventional pathway for exosome generation.
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影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.1126/science.aba2894
发表时间：
2020-10-16
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Lomakin AJ;Cattin CJ;Cuvelier D;Alraies Z;Molina M;Nader GPF;Srivastava N;Sáez PJ;Garcia-Arcos JM;Zhitnyak IY;Bhargava A;Driscoll MK;Welf ES;Fiolka R;Petrie RJ;De Silva NS;González-Granado JM;Manel N;Lennon-Duménil AM;Müller DJ;Piel M
通讯作者：
Piel M
影响因子：
14.2
作者：
Esser, Julia;Gehrmann, Ulf;Radmark, Olof
通讯作者：
Radmark, Olof
DOI：
10.1007/978-1-0716-0154-9_3
发表时间：
2020-01-01
期刊：
NEUTROPHIL: METHODS AND PROTOCOLS, 3RD EDITION
影响因子：
--
作者：
Kremserova, Silvie;Nauseef, William M.
通讯作者：
Nauseef, William M.
影响因子：
9.8
作者：
Majumdar R;Tavakoli Tameh A;Arya SB;Parent CA
通讯作者：
Parent CA