Apical localization of the Na-K-Cl cotransporter, rBSC1, on rat thick ascending limbs

Mark R. Kaplan, Matthew D. Plotkin, Wen Sen Lee, Zheng Chun Xu, Jonathan Lytton, Steven C. Hebert

Research output: Contribution to journalArticle

146 Citations (Scopus)

Abstract

A bumetanide-sensitive Na-K-Cl cotransporter (rBSC1) was recently cloned from a rat renal outer medulla (OM) cDNA library and shown to be expressed predominantly in the kidney [1]. The purpose of the present study was to examine the nephron distribution of cotransporter transcripts and protein in rat kidney. In situ hybridization showed an intense signal only in the outer medulla and extending along cortical medullary rays consistent with expression of rBSC1 transcripts in medullary (MTAL) and cortical (CTAL) thick ascending limbs. Polyclonal antibodies raised in rabbits against a unique 67 amino acid segment from the carboxyl terminus of rBSC1 identified a broad major band of 130 to 160 (midpoint of 150) kDa and at least two minor bands of 50 to 70 kD on Western blotting of homogenates from cortex (C) and outer medulla (OM), but not inner medulla (IM), of rat kidney. Thus the Na-K-Cl cotransporter protein detected by the polyclonal rBSC1 antibody in rat kidney was similar in size to the major ~150 kD bumetanide binding protein detected by others in mouse and dog kidneys. Immunofluorescence studies using the anti-rBSC1 polyclonal antibody on rat kidney sections showed an intense signal limited to apical surfaces of MTAL and CTAL segments. Colocalization with anti-Tamm-Horsfall antibody which is present in all TAL cells except macula densa cells confirmed the absence of anti-rBSC1 fluorescence in the macula densa cells. These results are consistent with rBSC1 encoding the, or the major isoform of the, apical Na-K-Cl cotransporter in the thick ascending limb. The Na-K-Cl cotransporter functionally detected in macula densa cells may be encoded by a different BSC isoform.

Original languageEnglish
Pages (from-to)40-47
Number of pages8
JournalKidney International
Volume49
Issue number1
Publication statusPublished - 1996
Externally publishedYes

Fingerprint

Sodium-Potassium-Chloride Symporters
Extremities
Kidney
Bumetanide
Antibodies
Protein Isoforms
Kidney Medulla
Nephrons
Gene Library
In Situ Hybridization
Fluorescent Antibody Technique
Carrier Proteins
Proteins
Fluorescence
Western Blotting
Dogs
Rabbits
Amino Acids

ASJC Scopus subject areas

  • Nephrology

Cite this

Kaplan, M. R., Plotkin, M. D., Lee, W. S., Xu, Z. C., Lytton, J., & Hebert, S. C. (1996). Apical localization of the Na-K-Cl cotransporter, rBSC1, on rat thick ascending limbs. Kidney International, 49(1), 40-47.

Apical localization of the Na-K-Cl cotransporter, rBSC1, on rat thick ascending limbs. / Kaplan, Mark R.; Plotkin, Matthew D.; Lee, Wen Sen; Xu, Zheng Chun; Lytton, Jonathan; Hebert, Steven C.

In: Kidney International, Vol. 49, No. 1, 1996, p. 40-47.

Research output: Contribution to journalArticle

Kaplan, MR, Plotkin, MD, Lee, WS, Xu, ZC, Lytton, J & Hebert, SC 1996, 'Apical localization of the Na-K-Cl cotransporter, rBSC1, on rat thick ascending limbs', Kidney International, vol. 49, no. 1, pp. 40-47.
Kaplan, Mark R. ; Plotkin, Matthew D. ; Lee, Wen Sen ; Xu, Zheng Chun ; Lytton, Jonathan ; Hebert, Steven C. / Apical localization of the Na-K-Cl cotransporter, rBSC1, on rat thick ascending limbs. In: Kidney International. 1996 ; Vol. 49, No. 1. pp. 40-47.
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