Water channel proteins in rat cardiac myocyte caveolae: osmolarity-dependent reversible internalization.
We show by confocal immunofluorescence microscopy that the water channel protein aquaporin-1, not previously identified within cardiomyocytes, localizes at 20 and 37 degrees C to rat cardiomyocyte sarcolemmal caveolar membrane and subsarcolemmal cytoplasm of primary atrial myocyte cultures, dissociated atrial and ventricular myocytes, and in situ cardiomyocytes of atrial and ventricular frozen sections. Confocal immunofluorescence microscopy shows that the normal in situ colocalization of the quasi-muscle-specific caveolar coating protein caveolin-3 with aquaporin-1 is reversibly disrupted by exposing in situ atrial or ventricular myocytes to physiological saline made hypertonic by adding 150 mM sucrose or 75 mM NaCl to isotonic physiological saline. This causes caveolae to close off from the interstitium and swell, while aquaporin-1 is internalized reversibly. At 4 degrees C aquaporin-1 does not colocalize with caveolin-3. We suggest that 1) in vivo, under near-isotonic conditions, caveolae may alternate frequently between brief open and closed-off states; 2) aquaporin-1-caveolin-3 colocalization may be energy dependent; and 3) while closed off from the interstitium, each caveola transiently functions as an osmometer that experiences, monitors, and reacts to net water flow from or into the subcaveolar cytosol of the myocyte.
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Departments of Medicine, University of Chicago, Chicago, Illinois 60637, USA.
Winterfield
J
J
Goings
G
G
Bastawrous
A
A
Upshaw-Earley
J
J
D000818:Animals; D051398:Aquaporin 1; D020346:Aquaporins / Q000032:analysis* / Q000378:metabolism; D051244:Caveolin 3; D022461:Caveolins; D002478:Cells, Cultured; D005260:Female; D006325:Heart Atria / Q000737:chemistry / Q000166:cytology; D006352:Heart Ventricles / Q000737:chemistry / Q000166:cytology; D008565:Membrane Proteins / Q000032:analysis* / Q000378:metabolism; D018613:Microscopy, Confocal; D009124:Muscle Proteins / Q000032:analysis* / Q000378:metabolism; D009206:Myocardium / Q000737:chemistry* / Q000166:cytology; D009994:Osmolar Concentration; D051381:Rats; D017207:Rats, Sprague-Dawley; D012508:Sarcolemma / Q000737:chemistry / Q000378:metabolism
C496022:Aqp1 protein, rat; D020346:Aquaporins; C495549:Cav3 protein, rat; D051244:Caveolin 3; D022461:Caveolins; D008565:Membrane Proteins; D009124:Muscle Proteins; D051398:Aquaporin 1