Hyperosmotic stimuli induces recruitment of aquaporin-1 to plasma membrane in cultured rat peritoneal mesothelial cells.
Aquaporin-1 (AQP-1) has been reported to play an important role in peritoneal dialysis. To determine the precise mechanism involved, we used cultured rat peritoneal mesothelial cells (RPMCs) to examine the translocation of AQP-1 to the plasma membrane induced by hyperosmotic stimuli. Cultured RPMCs obtained from male Sprague-Dawley rats were incubated at room temperature in Dulbecco modified Eagle medium/F12 (DMEM/F12) with and without glucose or mannitol as the hyperosmotic stimulus. The plasma membrane was then extracted by the Percoll gradient method. Finally, the abundance AQP-1 molecules in the membrane fraction was determined by Western blot analysis. Significant enhancement of AQP-1 abundance (p < 0.05) was observed within 2.5 minutes of the addition of 5% glucose to the medium. The increase was sustained in its abundance through 15 minutes. Abundance of AQP-1 was also increased (p < 0.05) by the addition of 5% mannitol. These results suggest that hyperosmotic stimuli could generate increased AQP-1 abundance in the plasma membrane by translocation of AQP-1 protein from recycling endosomes or early endosomes to the plasma membrane, rather than by protein synthesis via newly expressed mRNA. The latter mechanism would be expected to take more time.
Kuboshima
S
S
Division of Nephrology and Hypertension, St. Marianna University School of Medicine, Kawasaki, Japan.
Ogimoto
G
G
Sakurada
T
T
Fujino
T
T
Sato
T
T
Yasuda
T
T
Maeba
T
T
Owada
S
S
Ishida
M
M
D000818:Animals; D051398:Aquaporin 1; D020346:Aquaporins / Q000378:metabolism*; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003470:Culture Media; D004847:Epithelial Cells / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology; D008297:Male; D008353:Mannitol / Q000494:pharmacology*; D009994:Osmolar Concentration; D010537:Peritoneum / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
C496022:Aqp1 protein, rat; D020346:Aquaporins; D003470:Culture Media; D051398:Aquaporin 1; D008353:Mannitol; D005947:Glucose