Expression and subcellular localization of the AQP8 and AQP1 water channels in the mouse gall-bladder epithelium.
BACKGROUND
Transepithelial transport of water is one of the most distinctive functions by which the gall-bladder rearranges its bile content. Water is reabsorbed from the gall-bladder lumen during fasting, whereas it is secreted into the lumen following meal ingestion. Nevertheless, the molecular mechanism by which water is transported across the gall-bladder epithelium remains mostly unclear.


RESULTS
In the present study, we investigate the presence and subcellular localization of AQP (aquaporin) water channels in the mouse gall-bladder epithelium. Considerable AQP8 mRNA was detected in the gall-bladder epithelium of mouse, calf, rabbit, guinea pig and man. Studies of subcellular localization were then addressed to the mouse gall-bladder where the transcript of a second AQP, AQP1, was also detected. Immunoblotting experiments confirmed the presence of AQP8 and AQP1 at a protein level. Immunohistochemistry showed intense expression of AQP8 and AQP1 in the gall-bladder epithelial cells where AQP8 was localized in the apical membrane, whereas AQP1 was seen both in the apical and basolateral membranes, and in vesicles located in the subapical cytoplasm.


CONCLUSIONS
The pattern of subcellular distribution of AQP8 and AQP1 strongly corroborates the hypothesis of a transcellular route for the movement of water across the gall-bladder epithelium. Osmotic water would cross the apical membrane through AQP8 and AQP1, although AQP1 would be the facilitated pathway for the movement of water across the basolateral membrane. The presence of two distinct AQPs in the apical membrane is an unusual finding and may relate to the membrane's ability both to absorb and secrete fluid. It is tempting to hypothesize that AQP1 is hormonally translocated to the gall-bladder apical membrane to secrete water as in the bile duct epithelium, a functional homologue of the gall-bladder epithelium, whereas apical AQP8 may account for the absorption of water from gall-bladder bile.
Calamita
Giuseppe
G
Department of General and Environmental Physiology, University of Bari, via Amendola, 165/A-I-70126 Bari, Italy. calamita@biologia.uniba.it
Ferri
Domenico
D
Bazzini
Claudia
C
Mazzone
Amelia
A
Bottà
Guido
G
Liquori
Giuseppa E
GE
Paulmichl
Markus
M
Portincasa
Piero
P
Meyer
Giuliano
G
Svelto
M
M
D000595:Amino Acid Sequence; D000818:Animals; D051398:Aquaporin 1; D020346:Aquaporins / Q000096:biosynthesis* / Q000737:chemistry; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism*; D005704:Gallbladder / Q000378:metabolism*; D005786:Gene Expression Regulation; D006168:Guinea Pigs; D006728:Hormones / Q000378:metabolism; D015151:Immunoblotting; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007473:Ion Channels / Q000096:biosynthesis* / Q000737:chemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014867:Water / Q000378:metabolism
C496021:Aqp1 protein, mouse; D020346:Aquaporins; D006728:Hormones; D007473:Ion Channels; D012333:RNA, Messenger; C108330:aquaporin 8; D014867:Water; D051398:Aquaporin 1