Investigating Mast Cell Secretory Granules; from Biosynthesis to Exocytosis

Nurit P. Azouz; Mitsunori Fukuda; Marc E. Rothenberg; Ronit Sagi-Eisenberg

doi:10.3791/52505

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Investigating Mast Cell Secretory Granules; from Biosynthesis to Exocytosis

调查肥大细胞分泌颗粒;从合成到胞吐作用

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16:01 min

January 26, 2015

DOI: 10.3791/52505-v

Nurit P. Azouz^1,2, Mitsunori Fukuda³, Marc E. Rothenberg², Ronit Sagi-Eisenberg¹

¹Department of Cell and Developmental Biology,Sackler Faculty of Medicine, Tel Aviv University, ²Division of Allergy and Immunology, Department of Pediatrics,Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, ³Laboratory of Membrane Trafficking Mechanisms, Department of Developmental Biology and Neurosciences,Graduate School of Life Sciences, Tohoku University

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本议定书的目标是开发一种方法，使肥大细胞分泌功能基因组分析。所述协议是基于荧光报道基因cotrasfected与感兴趣和实时的基因的释放定量评估分析分泌颗粒的形态。

该程序的总体目标是研究调节肥大细胞、分泌颗粒发育和胞细胞作用的机制。这是通过首先用胞吐作用的报告基因以及目标基因横切肥大细胞来实现的。第二步是通过刺激细胞和监测胞吐过程中报告基因的荧光信号来评估测试基因对胞吐作用的影响。

接下来，通过定位于秘书颗粒的报告基因的共聚焦成像来评估测试基因对肥大细胞秘书颗粒生成和发育的影响。最后一步是对分泌颗粒进行形态计量分析。最终，使用报告基因对肥大细胞分泌颗粒进行荧光标记用于显示肥大细胞的改变、分泌颗粒发育及其胞吐作用。

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免疫学第95 肥大细胞胞外分泌分泌颗粒活细胞成像 Rab5的报告基因共焦成像转