A primary culture of distal convoluted tubules expressing functional thiazide-sensitive NaCl transport
Fulltext:
107770.pdf
Embargo:
until further notice
Size:
794.4Kb
Format:
PDF
Description:
Publisher’s version
Publication year
2012Source
American Journal of Physiology : Renal Physiology, 303, 6, (2012), pp. F886-92ISSN
Publication type
Article / Letter to editor
Display more detailsDisplay less details
Organization
Physiology
Journal title
American Journal of Physiology : Renal Physiology
Volume
vol. 303
Issue
iss. 6
Page start
p. F886
Page end
p. 92
Subject
NCMLS 5: Membrane transport and intracellular motility IGMD 9: Renal disorderAbstract
Studying the molecular regulation of the thiazide-sensitive Na(+)-Cl(-) cotransporter (NCC) is important for understanding how the kidney contributes to blood pressure regulation. Until now, a native mammalian cell model to investigate this transporter remained unknown. Our aim here is to establish, for the first time, a primary distal convoluted tubule (DCT) cell culture exhibiting transcellular thiazide-sensitive Na(+) transport. Because parvalbumin (PV) is primarily expressed in the DCT, where it colocalizes with NCC, kidneys from mice expressing enhanced green-fluorescent protein (eGFP) under the PV gene promoter (PV-eGFP-mice) were employed. The Complex Object Parametric Analyzer and Sorter (COPAS) was used to sort fluorescent PV-positive tubules from these kidneys, which were then seeded onto permeable supports. After 6 days, DCT cell monolayers developed transepithelial resistance values of 630 +/- 33 Omega.cm(2). The monolayers also established opposing transcellular concentration gradients of Na(+) and K(+). Radioactive (22)Na(+) flux experiments showed a net apical-to-basolateral thiazide-sensitive Na(+) transport across the monolayers. Both hypotonic low-chloride medium and 1 muM angiotensin II increased this (22)Na(+) transport significantly by four times, which could be totally blocked by 100 muM hydrochlorothiazide. Angiotensin II-stimulated (22)Na(+) transport was also inhibited by 1 muM losartan. Furthermore, NCC present in the DCT monolayers was detected by immunoblot and immunocytochemistry studies. In conclusion, a murine primary DCT culture was established which expresses functional thiazide-sensitive Na(+)-Cl(-) transport.
This item appears in the following Collection(s)
- Academic publications [238441]
- Electronic publications [122539]
- Faculty of Medical Sciences [90373]
Upload full text
Use your RU credentials (u/z-number and password) to log in with SURFconext to upload a file for processing by the repository team.