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Contribution of Aquaporins to Cellular Water Transport Observed by a Microfluidic Cell Volume Sensor
Here we demonstrate that an impedance-based microfluidic cell volume sensor can be used to study the roles of aquaporin (AQP) in cellular water permeability and screen AQP-specific drugs. Human embryonic kidney (HEK-293) cells were transiently transfected with AQP3- or AQP4-encoding genes to express...
Autores principales: | , , |
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Formato: | Texto |
Lenguaje: | English |
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American Chemical Society
2008
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2612540/ https://www.ncbi.nlm.nih.gov/pubmed/18698799 http://dx.doi.org/10.1021/ac8008498 |
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author | Heo, Jinseok Meng, Fanjie Hua, Susan Z. |
author_facet | Heo, Jinseok Meng, Fanjie Hua, Susan Z. |
author_sort | Heo, Jinseok |
collection | PubMed |
description | Here we demonstrate that an impedance-based microfluidic cell volume sensor can be used to study the roles of aquaporin (AQP) in cellular water permeability and screen AQP-specific drugs. Human embryonic kidney (HEK-293) cells were transiently transfected with AQP3- or AQP4-encoding genes to express AQPs in plasma membranes. The swelling of cells in response to hypotonic stimulation was traced in real time using the sensor. Two time constants were obtained by fitting the swelling curves with a two-exponential function, a fast time constant associated with osmotic water permeability of AQP-expressing cells and a slow phase time constant associated mainly with water diffusion through lipid bilayers in the nontransfected cells. The AQP-expressing cells showed at least 10× faster osmotic water transport than control cells. Using the volume sensor, we examined the effects of Hg(2+) and Ni(2+) on the water transport via AQPs. Hg(2+) inhibited the water flux in AQP3-expressing cells irreversibly, while Ni(2+) blocked the AQP3 channels reversibly. Neither of the two ions blocked the AQP4 channels. The microfluidic volume sensor can sense changes in cell volume in real time, which enables perfusion of various reagents sequentially. It provides a convenient tool for studying the effect of reagents on the function and regulation mechanism of AQPs. |
format | Text |
id | pubmed-2612540 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-26125402009-03-20 Contribution of Aquaporins to Cellular Water Transport Observed by a Microfluidic Cell Volume Sensor Heo, Jinseok Meng, Fanjie Hua, Susan Z. Anal Chem Here we demonstrate that an impedance-based microfluidic cell volume sensor can be used to study the roles of aquaporin (AQP) in cellular water permeability and screen AQP-specific drugs. Human embryonic kidney (HEK-293) cells were transiently transfected with AQP3- or AQP4-encoding genes to express AQPs in plasma membranes. The swelling of cells in response to hypotonic stimulation was traced in real time using the sensor. Two time constants were obtained by fitting the swelling curves with a two-exponential function, a fast time constant associated with osmotic water permeability of AQP-expressing cells and a slow phase time constant associated mainly with water diffusion through lipid bilayers in the nontransfected cells. The AQP-expressing cells showed at least 10× faster osmotic water transport than control cells. Using the volume sensor, we examined the effects of Hg(2+) and Ni(2+) on the water transport via AQPs. Hg(2+) inhibited the water flux in AQP3-expressing cells irreversibly, while Ni(2+) blocked the AQP3 channels reversibly. Neither of the two ions blocked the AQP4 channels. The microfluidic volume sensor can sense changes in cell volume in real time, which enables perfusion of various reagents sequentially. It provides a convenient tool for studying the effect of reagents on the function and regulation mechanism of AQPs. American Chemical Society 2008-08-13 2008-09-15 /pmc/articles/PMC2612540/ /pubmed/18698799 http://dx.doi.org/10.1021/ac8008498 Text en Copyright © 2008 American Chemical Society http://pubs.acs.org This is an open-access article distributed under the ACS AuthorChoice Terms & Conditions. Any use of this article, must conform to the terms of that license which are available at http://pubs.acs.org. 40.75 |
spellingShingle | Heo, Jinseok Meng, Fanjie Hua, Susan Z. Contribution of Aquaporins to Cellular Water Transport Observed by a Microfluidic Cell Volume Sensor |
title | Contribution of Aquaporins to Cellular Water Transport Observed by a Microfluidic Cell Volume Sensor |
title_full | Contribution of Aquaporins to Cellular Water Transport Observed by a Microfluidic Cell Volume Sensor |
title_fullStr | Contribution of Aquaporins to Cellular Water Transport Observed by a Microfluidic Cell Volume Sensor |
title_full_unstemmed | Contribution of Aquaporins to Cellular Water Transport Observed by a Microfluidic Cell Volume Sensor |
title_short | Contribution of Aquaporins to Cellular Water Transport Observed by a Microfluidic Cell Volume Sensor |
title_sort | contribution of aquaporins to cellular water transport observed by a microfluidic cell volume sensor |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2612540/ https://www.ncbi.nlm.nih.gov/pubmed/18698799 http://dx.doi.org/10.1021/ac8008498 |
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