Both Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>

As sessile organisms, plants must directly deal with an often complex and adverse environment in which hyperosmotic stress is one of the most serious abiotic factors, challenging cellular physiology and integrity. The plasma membrane (PM) is the hydrophobic barrier between the inside and outside env...

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Autores principales:	Zheng Wu, Chengyu Fan, Yi Man, Yue Zhang, Ruili Li, Xiaojuan Li, Yanping Jing
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	endocytosis clathrin microdomain hyperosmotic stress <i>Arabidopsis thaliana</i> Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/82a6a629d0294101979dd4d8711f5492
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spelling	oai:doaj.org-article:82a6a629d0294101979dd4d8711f54922021-11-25T17:57:35ZBoth Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>10.3390/ijms2222125341422-00671661-6596https://doaj.org/article/82a6a629d0294101979dd4d8711f54922021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12534https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067As sessile organisms, plants must directly deal with an often complex and adverse environment in which hyperosmotic stress is one of the most serious abiotic factors, challenging cellular physiology and integrity. The plasma membrane (PM) is the hydrophobic barrier between the inside and outside environments of cells and is considered a central compartment in cellular adaptation to diverse stress conditions through dynamic PM remodeling. Endocytosis is a powerful method for rapid remodeling of the PM. In animal cells, different endocytic pathways are activated in response to osmotic stress, while only a few reports are related to the endocytosis response pathway and involve a mechanism in plant cells upon hyperosmotic stress. In this study, using different endocytosis inhibitors, the microdomain-specific dye di-4-ANEPPDHQ, variable-angle total internal reflection fluorescence microscopy (VA-TIRFM), and confocal microscopy, we discovered that internalized Clathrin Light Chain-Green Fluorescent Protein (CLC-GFP) increased under hyperosmotic conditions, accompanied by decreased fluorescence intensity of CLC-GFP at the PM. CLC-GFP tended to have higher diffusion coefficients and a fraction of CLC-GFP molecules underwent slower diffusion upon hyperosmotic stress. Meanwhile, an increased motion range of CLC-GFP was found under hyperosmotic treatment compared with the control. In addition, the order of the PM decreased, but the order of the endosome increased when cells were in hyperosmotic conditions. Hence, our results demonstrated that clathrin-mediated endocytosis and membrane microdomain-associated endocytosis both participate in the adaptation to hyperosmotic stress. These findings will help to further understand the role and the regulatory mechanism involved in plant endocytosis in helping plants adapt to osmotic stress.Zheng WuChengyu FanYi ManYue ZhangRuili LiXiaojuan LiYanping JingMDPI AGarticleendocytosisclathrinmicrodomainhyperosmotic stress<i>Arabidopsis thaliana</i>Biology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12534, p 12534 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	endocytosis clathrin microdomain hyperosmotic stress <i>Arabidopsis thaliana</i> Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	endocytosis clathrin microdomain hyperosmotic stress <i>Arabidopsis thaliana</i> Biology (General) QH301-705.5 Chemistry QD1-999 Zheng Wu Chengyu Fan Yi Man Yue Zhang Ruili Li Xiaojuan Li Yanping Jing Both Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>
description	As sessile organisms, plants must directly deal with an often complex and adverse environment in which hyperosmotic stress is one of the most serious abiotic factors, challenging cellular physiology and integrity. The plasma membrane (PM) is the hydrophobic barrier between the inside and outside environments of cells and is considered a central compartment in cellular adaptation to diverse stress conditions through dynamic PM remodeling. Endocytosis is a powerful method for rapid remodeling of the PM. In animal cells, different endocytic pathways are activated in response to osmotic stress, while only a few reports are related to the endocytosis response pathway and involve a mechanism in plant cells upon hyperosmotic stress. In this study, using different endocytosis inhibitors, the microdomain-specific dye di-4-ANEPPDHQ, variable-angle total internal reflection fluorescence microscopy (VA-TIRFM), and confocal microscopy, we discovered that internalized Clathrin Light Chain-Green Fluorescent Protein (CLC-GFP) increased under hyperosmotic conditions, accompanied by decreased fluorescence intensity of CLC-GFP at the PM. CLC-GFP tended to have higher diffusion coefficients and a fraction of CLC-GFP molecules underwent slower diffusion upon hyperosmotic stress. Meanwhile, an increased motion range of CLC-GFP was found under hyperosmotic treatment compared with the control. In addition, the order of the PM decreased, but the order of the endosome increased when cells were in hyperosmotic conditions. Hence, our results demonstrated that clathrin-mediated endocytosis and membrane microdomain-associated endocytosis both participate in the adaptation to hyperosmotic stress. These findings will help to further understand the role and the regulatory mechanism involved in plant endocytosis in helping plants adapt to osmotic stress.
format	article
author	Zheng Wu Chengyu Fan Yi Man Yue Zhang Ruili Li Xiaojuan Li Yanping Jing
author_facet	Zheng Wu Chengyu Fan Yi Man Yue Zhang Ruili Li Xiaojuan Li Yanping Jing
author_sort	Zheng Wu
title	Both Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>
title_short	Both Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>
title_full	Both Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>
title_fullStr	Both Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>
title_full_unstemmed	Both Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>
title_sort	both clathrin-mediated and membrane microdomain-associated endocytosis contribute to the cellular adaptation to hyperosmotic stress in <i>arabidopsis</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/82a6a629d0294101979dd4d8711f5492
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Both Clathrin-Mediated and Membrane Microdomain-Associated Endocytosis Contribute to the Cellular Adaptation to Hyperosmotic Stress in <i>Arabidopsis</i>

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