NDLI: A bumetanide-sensitive, apically localized Na+2Cl–K+ cotransport in the rat inner medullary collecting duct

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A bumetanide-sensitive, apically localized Na+2Cl–K+ cotransport in the rat inner medullary collecting duct

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A bumetanide-sensitive, apically localized Na+2Cl–K+ cotransport in the rat inner medullary collecting duct

Content Provider	Springer Nature Link
Author	Grupp, C. Stelmach, R. Troche, I. Müller, G.
Copyright Year	1999
Abstract	To better characterize loop diuretic-sensitive ion fluxes in the inner medullary collecting duct (IMCD) we examined them in IMCD cells grown as a primary culture on permeable supports. A polarization of the cells with their basolateral side to the support was confirmed morphologically by electron microscopy and functionally by flux studies with ouabain. Within 7 days cells developed a transepithelial resistance of 974±52 Ω per cm2 and a low transepithelial potential difference (–0.7±0.8 mV). Measurements of intracellular ion content by electron probe microanalysis in IMCD depleted of intracellular ions by preincubation in a Na+-K+-Cl–-free medium revealed, compared to the control receiving solvent, significant reductions in intracellular Na+ content (–17.6% within 10 min) and intracellular Cl– content (–43.8% within 30 min) by the addition of bumetanide (10–4 mol/l) to the apical but not basolateral incubation medium. In 22Na+ and 86Rb+ isotope uptake studies, fluxes from the apical side were significantly inhibited at bumetanide concentrations of 100 µmol/l by 0.27±0.10 and 0.21±0.04 nmol/cm2 in 10 min, respectively, whereas basolateral fluxes of 86Rb+ but not 22Na+ were significantly reduced by this substance. Removal of Cl– had a similar but not additional effect. mRNA encoding the apical isoform of the Na+2Cl–K+ cotransporter could be specifically amplified by reverse transcriptase polymerase chain reaction from the inner medulla and highly purified IMCD cells. Northern blot of mRNA isolated from the inner medulla with a riboprobe of the apical isoform revealed a transcript of ~ 4.9 kb. This probe localized under "low-stringency" conditions to the IMCD in in situ hybridization studies. These results suggest the presence of an apically localized isoform of bumetanide-sensitive Na+2Cl–K+ cotransport in at least a subfraction of IMCD cells. This transport may be involved in the ultimate adjustment of urinary electrolyte concentration by this final segment of the tubular system.
Starting Page	174
Ending Page	185
Page Count	12
File Format	PDF
ISSN	00316768
Journal	Pflügers Archiv
Volume Number	439
Issue Number	1-2
e-ISSN	14322013
Language	English
Publisher	Springer-Verlag
Publisher Date	1999-09-28
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Content Type	Text
Resource Type	Article
Subject	Physiology Physiology (medical) Clinical Biochemistry

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