A new model of the distal convoluted tubule

被引:38
作者
Ko, Benjamin [1 ]
Mistry, Abinash C. [2 ]
Hanson, Lauren [1 ]
Mallick, Rickta [2 ]
Cooke, Leslie L. [1 ]
Hack, Bradley K. [1 ]
Cunningham, Patrick [1 ]
Hoover, Robert S. [2 ,3 ]
机构
[1] Univ Chicago, Dept Med, Chicago, IL 60637 USA
[2] Emory Univ, Dept Med, Div Nephrol, Atlanta, GA 30322 USA
[3] Atlanta Vet Adm Med Ctr, Decatur, GA USA
关键词
sodium-chloride cotransporter; STE20/SPS-1-related proline/alanine-rich kinase; with-no-lysine kinase 4; SENSITIVE NACL COTRANSPORTER; NA+-CL-COTRANSPORTER; SODIUM-CHLORIDE COTRANSPORTER; MOLECULAR PATHOGENESIS; SURFACE EXPRESSION; STIMULATION; MECHANISM; TRANSPORT; CHANNELS; KINASES;
D O I
10.1152/ajprenal.00139.2012
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
Ko B, Mistry AC, Hanson L, Mallick R, Cooke LL, Hack BK, Cunningham P, Hoover RS. A new model of the distal convoluted tubule. Am J Physiol Renal Physiol 303: F700-F710, 2012. First published June 20, 2012; doi: 10.1152/ajprenal.00139.2012.-The Na+-Cl- cotransporter (NCC) in the distal convoluted tubule (DCT) of the kidney is a key determinant of Na+ balance. Disturbances in NCC function are characterized by disordered volume and blood pressure regulation. However, many details concerning the mechanisms of NCC regulation remain controversial or undefined. This is partially due to the lack of a mammalian cell model of the DCT that is amenable to functional assessment of NCC activity. Previously reported investigations of NCC regulation in mammalian cells have either not attempted measurements of NCC function or have required perturbation of the critical without a lysine kinase (WNK)/STE20/SPS-1-related proline/alanine-rich kinase regulatory pathway before functional assessment. Here, we present a new mammalian model of the DCT, the mouse DCT15 (mDCT15) cell line. These cells display native NCC function as measured by thiazide-sensitive, Cl--dependent Na-22(+) uptake and allow for the separate assessment of NCC surface expression and activity. Knockdown by short interfering RNA confirmed that this function was dependent on NCC protein. Similar to the mammalian DCT, these cells express many of the known regulators of NCC and display significant baseline activity and dimerization of NCC. As described in previous models, NCC activity is inhibited by appropriate concentrations of thiazides, and phorbol esters strongly suppress function. Importantly, they display release of WNK4 inhibition of NCC by small hairpin RNA knockdown. We feel that this new model represents a critical tool for the study of NCC physiology. The work that can be accomplished in such a system represents a significant step forward toward unraveling the complex regulation of NCC.
引用
收藏
页码:F700 / F710
页数:11
相关论文
共 41 条
  • [1] WNK4 kinase regulates surface expression of the human sodium chloride cotransporter in mammalian cells
    Cai, H.
    Cebotaru, V.
    Wang, J. -H.
    Zhang, X. -M.
    Cebotaru, L.
    Guggino, S. E.
    Guggino, W. B.
    [J]. KIDNEY INTERNATIONAL, 2006, 69 (12) : 2162 - 2170
  • [2] Dietary salt regulates the phosphorylation of OSR1/SPAK kinases and the sodium chloride cotransporter through aldosterone
    Chiga, Motoko
    Rai, Tatemitsu
    Yang, Sung-Sen
    Ohta, Akihito
    Takizawa, Toichiro
    Sasaki, Sei
    Uchida, Shinichi
    [J]. KIDNEY INTERNATIONAL, 2008, 74 (11) : 1403 - 1409
  • [3] Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure
    Chobanian, AV
    Bakris, GL
    Black, HR
    Cushman, WC
    Green, LA
    Izzo, JL
    Jones, DW
    Materson, BJ
    Oparil, S
    Wright, JT
    Roccella, EJ
    [J]. HYPERTENSION, 2003, 42 (06) : 1206 - 1252
  • [4] Effects of chemical chaperones on partially retarded NaCl cotransporter mutants associated with Gitelman's syndrome in a mouse cortical collecting duct cell line
    de Jong, JC
    Willems, PHGM
    Goossens, M
    Vandewalle, A
    van den Heuvel, LPWJ
    Knoers, NVAM
    Bindels, RJM
    [J]. NEPHROLOGY DIALYSIS TRANSPLANTATION, 2004, 19 (05) : 1069 - 1076
  • [5] Functional expression of the human thiazide-sensitive NaCl cotransporter in Madin-Darby canine kidney cells
    De Jong, JC
    Willems, PHGM
    Van den Heuvel, LPWJ
    Knoers, NVAM
    Bindels, RJM
    [J]. JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, 2003, 14 (10): : 2428 - 2435
  • [6] The structural unit of the thiazide-sensitive NaCl cotransporter is a homodimer
    de Jong, JC
    Willems, PHGM
    Mooren, FJM
    van den Heuvel, LPWJ
    Knoers, NVAM
    Bindels, RJM
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (27) : 24302 - 24307
  • [7] γ-Adducin Stimulates the Thiazide-sensitive NaCl Cotransporter
    Dimke, Henrik
    San-Cristobal, Pedro
    de Graaf, Mark
    Lenders, Jacques W.
    Deinum, Jaap
    Hoenderop, Joost G. J.
    Bindels, Rene J. M.
    [J]. JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, 2011, 22 (03): : 508 - 517
  • [8] THIAZIDE-SENSITIVE SODIUM-CHLORIDE COTRANSPORT IN EARLY DISTAL TUBULE
    ELLISON, DH
    VELAZQUEZ, H
    WRIGHT, FS
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY, 1987, 253 (03): : F546 - F554
  • [9] Na channels in the rat connecting tubule
    Frindt, G
    Palmer, LG
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY, 2004, 286 (04) : F669 - F674
  • [10] Surface expression of sodium channels and transporters in rat kidney: effects of dietary sodium
    Frindt, Gustavo
    Palmer, Lawrence G.
    [J]. AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY, 2009, 297 (05) : F1249 - F1255