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Expression of proximal tubular Na-Pi and Na-SO4 cotransporters in MDCK and LLC-PK1 cells by transfection.


Quabius, E S; Murer, H; Biber, J (1996). Expression of proximal tubular Na-Pi and Na-SO4 cotransporters in MDCK and LLC-PK1 cells by transfection. American Journal of Physiology: Renal Physiology, 270(1 Pt 2):F220-F228.

Abstract

Two cD-NAs coding for proximal tubular Na-Pi cotransport (NaPi-2) and Na-SO4 cotransport (NaSi-1) have been transfected by the use of a dexamethasone-inducible vector (pLK-neo) into MDCK and LLC-PK1 cells. By reverse transcription-polymerase chain reaction, expression of corresponding mRNAs was observed after stimulation with dexamethasone only. Similarly, expression of the NaPi-2 protein was detected only after induction with dexamethasone. In transfected Madin-Darby canine kidney (MDCK) cells, dexamethasone induced a large increase of Na-Pi or Na-SO4 cotransport, whereas, in transfected LLC-PK1, cell transport was only minimally expressed. In MDCK cells grown on filter supports, transfected Na-Pi-cotransport activity was equally expressed at both cell surfaces; dual location of expressed NaPi-2 protein was also observed by immunohistochemistry. In contrast, transfected Na-SO4 cotransport activity was predominantly expressed at the apical cell surface of MDCK cells. The results demonstrate that 1), in MDCK cells, the sorting behavior of two proximal tubular cotransport systems seems to be different: apical for Na-SO4 cotransport (NaSi-1) and dual location for Na-Pi cotransport (NaPi-2); and 2) LLC-PK1 cells seem not to be a suitable system to functionally express sodium-dependent cotransport systems for phosphate and sulfate.

Two cD-NAs coding for proximal tubular Na-Pi cotransport (NaPi-2) and Na-SO4 cotransport (NaSi-1) have been transfected by the use of a dexamethasone-inducible vector (pLK-neo) into MDCK and LLC-PK1 cells. By reverse transcription-polymerase chain reaction, expression of corresponding mRNAs was observed after stimulation with dexamethasone only. Similarly, expression of the NaPi-2 protein was detected only after induction with dexamethasone. In transfected Madin-Darby canine kidney (MDCK) cells, dexamethasone induced a large increase of Na-Pi or Na-SO4 cotransport, whereas, in transfected LLC-PK1, cell transport was only minimally expressed. In MDCK cells grown on filter supports, transfected Na-Pi-cotransport activity was equally expressed at both cell surfaces; dual location of expressed NaPi-2 protein was also observed by immunohistochemistry. In contrast, transfected Na-SO4 cotransport activity was predominantly expressed at the apical cell surface of MDCK cells. The results demonstrate that 1), in MDCK cells, the sorting behavior of two proximal tubular cotransport systems seems to be different: apical for Na-SO4 cotransport (NaSi-1) and dual location for Na-Pi cotransport (NaPi-2); and 2) LLC-PK1 cells seem not to be a suitable system to functionally express sodium-dependent cotransport systems for phosphate and sulfate.

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Additional indexing

Item Type:Journal Article, refereed
Communities & Collections:04 Faculty of Medicine > Institute of Physiology
07 Faculty of Science > Institute of Physiology
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:1 January 1996
Deposited On:11 Feb 2008 12:22
Last Modified:05 Apr 2016 12:18
Publisher:American Physiological Society
ISSN:0002-9513
Related URLs:http://ajprenal.physiology.org/cgi/content/abstract/270/1/F220
PubMed ID:8769843

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