通过活体显微镜检查探究肌动蛋白细胞骨架在调节性胞吐作用中的作用。
Probing the role of the actin cytoskeleton during regulated exocytosis by intravital microscopy.
作者信息
Milberg Oleg, Tora Muhibullah, Shitara Akiko, Takuma Taishin, Masedunskas Andrius, Weigert Roberto
机构信息
Intracellular Membrane Trafficking Unit, Oral and Pharyngeal Cancer Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, 30 Convent Dr. 303A, 20892-4340, Bethesda, MD, USA.
出版信息
Methods Mol Biol. 2014;1174:407-21. doi: 10.1007/978-1-4939-0944-5_28.
Abstract
The actin cytoskeleton plays a fundamental role in controlling several steps during regulated exocytosis. Here, we describe a combination of procedures that are aimed at studying the dynamics and the mechanism of the actin cytoskeleton in the salivary glands of live rodents, a model for exocrine secretion. Our approach relies on intravital microscopy, an imaging technique that enables imaging biological events in live animals at a subcellular resolution, and it is complemented by the use of pharmacological agents and indirect immunofluorescence in the salivary tissue.
摘要
肌动蛋白细胞骨架在调节性胞吐作用的多个步骤控制中发挥着基本作用。在此，我们描述了一系列方法的组合，旨在研究活啮齿动物唾液腺中肌动蛋白细胞骨架的动力学和机制，唾液腺是外分泌分泌的一个模型。我们的方法依赖于活体显微镜检查，这是一种能够以亚细胞分辨率对活体动物中的生物事件进行成像的技术，并辅以在唾液组织中使用药理试剂和间接免疫荧光。
相似文献
1
Probing the role of the actin cytoskeleton during regulated exocytosis by intravital microscopy.通过活体显微镜检查探究肌动蛋白细胞骨架在调节性胞吐作用中的作用。
Methods Mol Biol. 2014;1174:407-21. doi: 10.1007/978-1-4939-0944-5_28.
2
Concerted actions of distinct nonmuscle myosin II isoforms drive intracellular membrane remodeling in live animals.不同的非肌肉肌球蛋白II亚型的协同作用驱动活体动物细胞内膜重塑。
J Cell Biol. 2017 Jul 3;216(7):1925-1936. doi: 10.1083/jcb.201612126. Epub 2017 Jun 9.
3
Role for the actomyosin complex in regulated exocytosis revealed by intravital microscopy.肌球蛋白复合物在活细胞内显影介导的胞吐作用中的作用。
Proc Natl Acad Sci U S A. 2011 Aug 16;108(33):13552-7. doi: 10.1073/pnas.1016778108. Epub 2011 Aug 1.
4
Intravital microscopy for imaging subcellular structures in live mice expressing fluorescent proteins.用于对表达荧光蛋白的活体小鼠亚细胞结构进行成像的活体显微镜检查。
J Vis Exp. 2013 Sep 1(79):50558. doi: 10.3791/50558.
5
Swip-1 promotes exocytosis of glue granules in the exocrine Drosophila salivary gland.Swip-1 促进了果蝇唾腺细胞中胶粒的胞吐作用。
J Cell Sci. 2023 Mar 15;136(6). doi: 10.1242/jcs.260366. Epub 2023 Mar 5.
6
Measurement of dynamic F-actin changes during exocytosis.胞吐过程中动态F-肌动蛋白变化的测量。
Methods Mol Biol. 2014;1174:423-31. doi: 10.1007/978-1-4939-0944-5_29.
7
Multiple roles for the actin cytoskeleton during regulated exocytosis.肌动蛋白细胞骨架在调节胞吐作用中的多种作用。
Cell Mol Life Sci. 2013 Jun;70(12):2099-121. doi: 10.1007/s00018-012-1156-5. Epub 2012 Sep 18.
8
Arp2/3-mediated F-actin formation controls regulated exocytosis in vivo.Arp2/3介导的F-肌动蛋白形成在体内控制调节性胞吐作用。
Nat Commun. 2015 Dec 7;6:10098. doi: 10.1038/ncomms10098.
9
Real-time insights into regulated exocytosis.对调节性胞吐作用的实时洞察。
J Cell Sci. 2017 Apr 15;130(8):1355-1363. doi: 10.1242/jcs.193425. Epub 2017 Mar 16.
10
Regulated exocytosis: novel insights from intravital microscopy.受调控的胞吐作用：活体显微镜的新见解。
Traffic. 2012 May;13(5):627-34. doi: 10.1111/j.1600-0854.2012.01328.x. Epub 2012 Jan 31.
引用本文的文献
1
Cdc42 negatively regulates endocytosis during apical membrane maintenance in live animals.Cdc42 在活体动物中顶膜维持期间负调控内吞作用。
Mol Biol Cell. 2019 Feb 1;30(3):324-332. doi: 10.1091/mbc.E18-10-0615. Epub 2018 Dec 12.
2
Concerted actions of distinct nonmuscle myosin II isoforms drive intracellular membrane remodeling in live animals.不同的非肌肉肌球蛋白II亚型的协同作用驱动活体动物细胞内膜重塑。
J Cell Biol. 2017 Jul 3;216(7):1925-1936. doi: 10.1083/jcb.201612126. Epub 2017 Jun 9.
3
Live-cell imaging of ER-PM contact architecture by a novel TIRFM approach reveals extension of junctions in response to store-operated Ca-entry.通过一种新型全内反射荧光显微镜（TIRFM）方法对内质网-质膜（ER-PM）接触结构进行活细胞成像，揭示了连接在储存式钙内流响应中的延伸。
Sci Rep. 2016 Oct 19;6:35656. doi: 10.1038/srep35656.
4
Imaging membrane remodeling during regulated exocytosis in live mice.在活体小鼠的调节性胞吐过程中对膜重塑进行成像。
Exp Cell Res. 2015 Oct 1;337(2):219-25. doi: 10.1016/j.yexcr.2015.06.018. Epub 2015 Jul 6.
本文引用的文献
1
Intravital microscopy for imaging subcellular structures in live mice expressing fluorescent proteins.用于对表达荧光蛋白的活体小鼠亚细胞结构进行成像的活体显微镜检查。
J Vis Exp. 2013 Sep 1(79):50558. doi: 10.3791/50558.
2
Imaging cell biology in live animals: ready for prime time.在活体动物中进行细胞生物学成像：准备好进入黄金时代。
J Cell Biol. 2013 Jun 24;201(7):969-79. doi: 10.1083/jcb.201212130.
3
Intravital microscopy to image membrane trafficking in live rats.在活体大鼠中成像膜运输的活体显微镜检查。
Methods Mol Biol. 2013;931:153-67. doi: 10.1007/978-1-62703-056-4_9.
4
Multiple roles for the actin cytoskeleton during regulated exocytosis.肌动蛋白细胞骨架在调节胞吐作用中的多种作用。
Cell Mol Life Sci. 2013 Jun;70(12):2099-121. doi: 10.1007/s00018-012-1156-5. Epub 2012 Sep 18.
5
Regulated exocytosis: novel insights from intravital microscopy.受调控的胞吐作用：活体显微镜的新见解。
Traffic. 2012 May;13(5):627-34. doi: 10.1111/j.1600-0854.2012.01328.x. Epub 2012 Jan 31.
6
Intravital imaging.活体成像。
Cell. 2011 Nov 23;147(5):983-91. doi: 10.1016/j.cell.2011.11.004.
7
Role for the actomyosin complex in regulated exocytosis revealed by intravital microscopy.肌球蛋白复合物在活细胞内显影介导的胞吐作用中的作用。
Proc Natl Acad Sci U S A. 2011 Aug 16;108(33):13552-7. doi: 10.1073/pnas.1016778108. Epub 2011 Aug 1.
8
Lifeact mice for studying F-actin dynamics.用于研究F-肌动蛋白动力学的Lifeact小鼠。
Nat Methods. 2010 Mar;7(3):168-9. doi: 10.1038/nmeth0310-168.
9
Differential sorting of human parathyroid hormone after transduction of mouse and rat salivary glands.小鼠和大鼠唾液腺转导后人甲状旁腺激素的差异分选
Hum Gene Ther. 2008 Oct;19(10):1021-8. doi: 10.1089/hum.2008.079.
10
Intravital two-photon microscopy for studying the uptake and trafficking of fluorescently conjugated molecules in live rodents.用于研究活体啮齿动物中荧光共轭分子摄取和运输的活体双光子显微镜技术。
Traffic. 2008 Sep;9(10):1801-10. doi: 10.1111/j.1600-0854.2008.00798.x. Epub 2008 Jul 18.
Zotero 插件