Expression and immunolocalization of aquaporin water channels in rat exocrine pancreas.
Both the acinar and ductal cells of the pancreas secrete a near-isotonic fluid and may thus be sites of aquaporin (AQP) water channel expression. Northern blot analysis of mRNA from whole rat pancreas revealed high levels of AQP1 and AQP8 expression, whereas lower levels of AQP4 and AQP5 expression were just detectable by RT-PCR Southern blot analysis. Immunohistochemistry showed that AQP1 is localized in the microvasculature, whereas AQP8 is confined to the apical pole of the acinar cells. No labeling of acinar, ductal, or vascular tissue was detected with antibodies to AQP2-7. With immunoelectron microscopy, AQP8 labeling was observed not only at the apical membrane of the acinar cells but also among small intracellular vesicles in the subapical cytoplasm, suggesting that there may be regulated trafficking of AQP8 to the apical plasma membrane. To evaluate the contribution of AQPs to the membrane water permeability, video microscopy was used to measure the swelling of acinar cells in response to hypotonic stress. Osmotic water permeability was reduced by 90% following exposure to Hg(2+). Since AQP8 is confined to the apical membrane, the marked effect of Hg(2+) suggests that other water channels may be expressed in the basolateral membrane.
Hurley
P T
PT
School of Biological Sciences, University of Manchester, Manchester M13 9PT, United Kingdom.
Ferguson
C J
CJ
Kwon
T H
TH
Andersen
M L
ML
Norman
A G
AG
Steward
M C
MC
Nielsen
S
S
Case
R M
RM
D000465:Algorithms; D000818:Animals; D020346:Aquaporins / Q000187:drug effects / Q000378:metabolism*; D015152:Blotting, Northern; D015139:Blotting, Southern; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D007038:Hypotonic Solutions; D007150:Immunohistochemistry; D008297:Male; D017669:Mercury Compounds / Q000494:pharmacology; D016253:Microscopy, Immunoelectron; D010179:Pancreas / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism
D020346:Aquaporins; D007038:Hypotonic Solutions; D017669:Mercury Compounds; D012313:RNA