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 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 10x 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.
Heo
Jinseok
J
Department of Mechanical & Aerospace Engineering, SUNY-Buffalo, Buffalo, New York 14260, USA.
Meng
Fanjie
F
Hua
Susan Z
SZ
D051400:Aquaporin 3 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051401:Aquaporin 4 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020346:Aquaporins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D048429:Cell Size / Q000187:drug effects; D002477:Cells / Q000166:cytology* / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007202:Indicators and Reagents / Q000494:pharmacology; D007700:Kinetics; D019216:Metals, Heavy / Q000494:pharmacology; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D010539:Permeability; D013379:Substrate Specificity; D014162:Transfection; D014867:Water / Q000378:metabolism*
D051401:Aquaporin 4; D020346:Aquaporins; D007202:Indicators and Reagents; D019216:Metals, Heavy; D014867:Water; D051400:Aquaporin 3