Am J Physiol Cell Physiol AJP: Renal Physiology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
QUICK SEARCH:   [advanced]


     

Am J Physiol Cell Physiol (January 19, 2005). doi:10.1152/ajpcell.00084.2004
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
288/5/C1134    most recent
00084.2004v2
00084.2004v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Web of Science (16)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Miller, R. L.
Right arrow Articles by Nelson, R. D
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Miller, R. L.
Right arrow Articles by Nelson, R. D
Submitted on February 10, 2004
Accepted on December 13, 2004

V-ATPase B1 Subunit Promoter Drives Expression Of EGFP In Intercalated Cells Of Kidney, Clear Cells Of Epididymis and Airway Cells of Lung In Transgenic Mice

R. Lance Miller, Ping Zhang, Maren Smith, Valerie Beaulieu, Teodor G Paunescu, Dennis Brown, Sylvie Breton, and Raoul D Nelson*

* To whom correspondence should be addressed. E-mail: Raoul.Nelson{at}hsc.utah.edu.

The kidney, epididymis and lungs are complex organs composed of tubules with considerable epithelial cell heterogeneity. This has limited the characterization of (patho)physiological transport processes that are specific for each cell type in these epithelia. The purpose of the present study was to develop new tools to study cell-specific gene and protein expression in such complex tissues and organs. Here, we report the production of a transgenic mouse that expresses the enhanced green fluorescent protein (EGFP) in a sub-set of epithelial cells, including intercalated cells in the kidney, and narrow and clear cells in the epididymis. These cells express the B1 subunit of the vacuolar H+ATPase (V-ATPase) and are actively involved in proton transport. A 6.5 kb portion of the V-ATPase B1 promoter was used to drive expression of EGFP. In two founders, quantitative real-time RT-PCR demonstrated expression of EGFP in kidney, epididymis and lung, but not in heart, brain, muscle, and liver. Immunofluorescence labeling using antibodies against the B1 and E subunits of the V-ATPase and against carbonic anhydrase type II (CAII) revealed specific EGFP expression in all renal type A and type B intercalated cells, some renal connecting tubule cells, all epididymal narrow and clear cells and some non-ciliated airway epithelial cells. No EGFP expression was detected in collecting duct principal cells (identified using an anti-AQP2 antibody) or epididymal principal cells (negative for V-ATPase or CAII). Thus, 6.5 kb of the V-ATPase B1 promoter is sufficient to confer cell specific expression. This EGFP expressing mouse model should prove useful in future studies of gene and protein expression and their physiological and/or developmental regulation in distinct cell types that can now be separated by fluorescence assisted microdissection, fluorescence activated cell sorting and laser capture microdissection.




This article has been cited by other articles:


Home page
 J. Exp. Biol.Home page
W. W. C. Shum, N. Da Silva, D. Brown, and S. Breton
Regulation of luminal acidification in the male reproductive tract via cell-cell crosstalk
J. Exp. Biol., June 1, 2009; 212(11): 1753 - 1761.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
T. G. Paunescu, A. C. Jones, R. Tyszkowski, and D. Brown
V-ATPase expression in the mouse olfactory epithelium
Am J Physiol Cell Physiol, October 1, 2008; 295(4): C923 - C930.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
T. G. Paunescu, L. M. Russo, N. Da Silva, J. Kovacikova, N. Mohebbi, A. N. Van Hoek, M. McKee, C. A. Wagner, S. Breton, and D. Brown
Compensatory membrane expression of the V-ATPase B2 subunit isoform in renal medullary intercalated cells of B1-deficient mice
Am J Physiol Renal Physiol, December 1, 2007; 293(6): F1915 - F1926.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
N. Da Silva, W. W. C. Shum, J. El-Annan, T. G. Paunescu, M. McKee, P. J. S. Smith, D. Brown, and S. Breton
Relocalization of the V-ATPase B2 subunit to the apical membrane of epididymal clear cells of mice deficient in the B1 subunit
Am J Physiol Cell Physiol, July 1, 2007; 293(1): C199 - C210.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
R. L. Miller, P. Zhang, T. Chen, A. Rohrwasser, and R. D. Nelson
Automated method for the isolation of collecting ducts
Am J Physiol Renal Physiol, July 1, 2006; 291(1): F236 - F245.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
R. A. Fenton, A. Shodeinde, and M. A. Knepper
UT-A urea transporter promoter, UT-A{alpha}, targets principal cells of the renal inner medullary collecting duct
Am J Physiol Renal Physiol, January 1, 2006; 290(1): F188 - F195.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
R. Sun, R. L. Miller, A. C. Hemmert, P. Zhang, H. Shi, R. D. Nelson, and B. K. Kishore
Chronic dDAVP infusion in rats decreases the expression of P2Y2 receptor in inner medulla and P2Y2 receptor-mediated PGE2 release by IMCD
Am J Physiol Renal Physiol, October 1, 2005; 289(4): F768 - F776.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Visit Other APS Journals Online
Copyright © 1977 by the American Physiological Society.