Identification and characterization of aquaporin-9 (AQP9) in porcine hepatic tissue and hepatocytes in monolayer culture.
Aquaporins (AQPs) are members of a large family of integral membrane proteins involved in the rapid movement of water and neutral solutes across cell membranes. In this study, we have prepared an affinity-purified porcine-specific polyclonal antiserum to AQP9 and have investigated the distribution and expression of AQP9 in pig liver tissue and in hepatocytes in primary culture. Immunocytochemical analysis showed that AQP9 was primarily localized in the membrane structures of hepatocytes and was not associated with intrahepatic bile ducts or blood vessels. Western blot analysis indicated that AQP9 ranged in apparent molecular mass between 27 and 38 kD in whole liver and hepatocyte membrane fractions; minor components were also observed at approximately 34 kD in the cytosol compartment of hepatocytes, bile duct and gall bladder. A prominent immunoreactive band at 44 kD was shown to be an artifact of Western blot analysis. In primary cultures of porcine hepatocytes, glucagon enhanced absolute levels of AQP9 protein, while gene expression was enhanced by T3 and glucagon. Insulin alone had no discernable influence on AQP9 gene expression or its cellular protein levels. These data suggest that AQP9 is a major AQP in porcine hepatic tissue and appears to be primarily responsive to glucagon induction.
Caperna
Thomas J
TJ
Growth Biology Laboratory, Animal and Natural Resources Institute, USDA/ARS, Beltsville Agricultural Research Center, USDA, Beltsville, MD 20705, United States. caperna@anri.barc.usda.gov
Shannon
Amy E
AE
Richards
Mark P
MP
Garrett
Wesley M
WM
Talbot
Neil C
NC
D000818:Animals; D020346:Aquaporins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D005934:Glucagon / Q000502:physiology; D022781:Hepatocytes / Q000378:metabolism*; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D034421:Sus scrofa; D013552:Swine
D020346:Aquaporins; D005934:Glucagon