Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1, in the rat kidney

Matthew D. Plotkin, Mark R. Kaplan, Jill W. Verlander, Wen Sen Lee, Dennis Brown, Esteban Poch, Steven R. Gullans, Steven C. Hebert

Research output: Contribution to journalArticle

157 Citations (Scopus)

Abstract

A thiazide sensitive Na-Cl contransporter, rTSC1, has recently been cloned from a rat kidney cortex cDNA library. The molecular regulation and nephron localization of this protein is unknown. The purpose of this study was to examine the nephron distribution and subcellular localization of the rTSC1 protein in the rat kidney. In situ hybridization showed rTSC1 transcripts were localized to short, convoluted tubule segments in the kidney cortex. Polyclonal antibodies raised against a 110 amino acid segment from the amino terminus of rTSC1 recognized three major bands of 135, 140 and 155 kDa on Western blotting of membrane protein from cortex but not outer medulla of the rat kidney. Immunofluorescence studies using the antibody alone and in double labeling experiments with antibodies against the H+ ATPase and calbindin D28, showed intense labeling of apical membranes in the distal nephron beginning in the initial distal convoluted tubule and terminating within the connecting tubule. The intensity of labeling diminished from proximal to distal sites along the distal tubule. Ultrastructural studies by immunoelectron microscopy showed the contransporter protein to be localized predominately on apical microvilli of the distal convoluted tubule cells. These results are consistent with rTSC1 encoding the apical thiazide sensitive Na-Cl cotransporter in the distal tubule.

Original languageEnglish
Pages (from-to)174-183
Number of pages10
JournalKidney International
Volume50
Issue number1
Publication statusPublished - 1996
Externally publishedYes

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Nephrons
Kidney Cortex
Kidney
Antibodies
Kidney Medulla
Thiazides
Calbindins
Proteins
Proton-Translocating ATPases
Immunoelectron Microscopy
Microvilli
Gene Library
In Situ Hybridization
Fluorescent Antibody Technique
Membrane Proteins
Western Blotting
Amino Acids
Membranes
thiazide receptor

ASJC Scopus subject areas

  • Nephrology

Cite this

Plotkin, M. D., Kaplan, M. R., Verlander, J. W., Lee, W. S., Brown, D., Poch, E., ... Hebert, S. C. (1996). Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1, in the rat kidney. Kidney International, 50(1), 174-183.

Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1, in the rat kidney. / Plotkin, Matthew D.; Kaplan, Mark R.; Verlander, Jill W.; Lee, Wen Sen; Brown, Dennis; Poch, Esteban; Gullans, Steven R.; Hebert, Steven C.

In: Kidney International, Vol. 50, No. 1, 1996, p. 174-183.

Research output: Contribution to journalArticle

Plotkin, MD, Kaplan, MR, Verlander, JW, Lee, WS, Brown, D, Poch, E, Gullans, SR & Hebert, SC 1996, 'Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1, in the rat kidney', Kidney International, vol. 50, no. 1, pp. 174-183.
Plotkin MD, Kaplan MR, Verlander JW, Lee WS, Brown D, Poch E et al. Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1, in the rat kidney. Kidney International. 1996;50(1):174-183.
Plotkin, Matthew D. ; Kaplan, Mark R. ; Verlander, Jill W. ; Lee, Wen Sen ; Brown, Dennis ; Poch, Esteban ; Gullans, Steven R. ; Hebert, Steven C. / Localization of the thiazide sensitive Na-Cl cotransporter, rTSC1, in the rat kidney. In: Kidney International. 1996 ; Vol. 50, No. 1. pp. 174-183.
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