|
|
||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
School of Biomedical Sciences, University of Leeds, Leeds LS2 9NQ, United Kingdom
AVP increases the osmotic water permeability of renal collecting ducts by inducing the translocation of specific aquaporin-2 (AQP2) water channels from cytoplasmic vesicles to the apical plasma membrane of the principal cells. Here, we report a novel inner medullary tubule suspension for the study of this phenomenon that overcomes some of the drawbacks faced by present techniques; both primary cultures of inner medullary collecting duct cells and cell lines expressing AQP2 show aberrant trafficking and/or signaling pathways. The tubule suspensions were prepared by proteolytic digestion of inner medullas dissected from freshly isolated rat kidneys. After drug treatment, cellular distribution of AQP2 was determined by membrane fractionation and Western blotting or by immunocytochemistry. Treatment of suspensions with 1 nM AVP caused redistribution of AQP2 to the apical plasma membrane of the principal cells, a process inhibited by microtubule disruption or PKA inhibition. We conclude that this method provides a valuable new approach to the study of the cellular mechanisms involved in the response of the collecting duct to AVP.
vesicle trafficking; new technique; collecting ducts; aquaporin-2
This article has been cited by other articles:
![]() |
K. Y. Renkema, A. Velic, H. B. Dijkman, S. Verkaart, A. W. van der Kemp, M. Nowik, K. Timmermans, A. Doucet, C. A. Wagner, R. J. Bindels, et al. The Calcium-Sensing Receptor Promotes Urinary Acidification to Prevent Nephrolithiasis J. Am. Soc. Nephrol., August 1, 2009; 20(8): 1705 - 1713. [Abstract] [Full Text] [PDF] |
||||
![]() |
S. Shaw and D. Marples N-ethylmaleimide causes aquaporin-2 trafficking in the renal inner medullary collecting duct by direct activation of protein kinase A Am J Physiol Renal Physiol, April 1, 2005; 288(4): F832 - F839. [Abstract] [Full Text] [PDF] |
||||
![]() |
J. Praetorius, Y.-H. Kim, E. V. Bouzinova, S. Frische, A. Rojek, C. Aalkjaer, and S. Nielsen NBCn1 is a basolateral Na+-HCO3 cotransporter in rat kidney inner medullary collecting ducts Am J Physiol Renal Physiol, May 1, 2004; 286(5): F903 - F912. [Abstract] [Full Text] [PDF] |
||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Visit Other APS Journals Online |