期刊
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY
卷 302, 期 5, 页码 F581-F590出版社
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajprenal.00304.2011
关键词
small GTPase; siRNA; dominant-negative Rab
资金
- National Institutes of Health (NIH) [K99DK078917, R37DK54425, R01DK077777, DK057718, DK047874, DK54814]
- Cystic Fibrosis Foundation [BUTTER06G0]
- Pittsburgh Center for Kidney Research [P30DK079307]
Butterworth MB, Edinger RS, Silvis MR, Gallo LI, Liang X, Apodaca G, Fizzell RA, Johnson JP. Rab11b regulates the trafficking and recycling of the epithelial sodium channel (ENaC). Am J Physiol Renal Physiol 302: F581-F590, 2012. First published November 30, 2011; doi: 10.1152/ajprenal.00304.2011.-Expression of the epithelial sodium channel (ENaC) at the apical membrane of cortical collecting duct (CCD) principal cells is modulated by regulated trafficking mediated by vesicle insertion and retrieval. Small GTPases are known to facilitate vesicle trafficking, recycling, and membrane fusion events; however, little is known about the specific Rab family members that modify ENaC surface density. Using a mouse CCD cell line that endogenously expresses ENaC (mpkCCD), the channel was localized to both Rab11a- and Rab11b-positive endosomes by immunoisolation and confocal fluorescent microscopy. Expression of a dominant negative (DN) form of Rab11a or Rab11b significantly reduced the basal and cAMP-stimulated ENaC-dependent sodium (Na+) transport. The greatest reduction in Na+ transport was observed with the expression of DN-Rab11b. Furthermore, small interfering RNA-mediated knockdown of each Rab11 isoform demonstrated the requirement for Rab11b in ENaC surface expression. These data indicate that Rab11b, and to a lesser extent Rab11a, is involved in establishing the constitutive and cAMP-stimulated Na+ transport in mpkCCD cells.
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