Cl- channels in basolateral TAL membranes: XIII. Heterogeneity between basolateral MTAL and CTAL Cl- channels

Christopher J. Winters, W. Brian Reeves, Thomas E. Andreoli

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Background. Antidiuretic hormone (ADH) or adenosine 3',5'-cyclic phosphate (cAMP) analogues augment net NaCl absorption in microperfused mouse medullary thick ascending limb (MTAL) segments but not in cortical thick ascending limb (CTAL) segments. This ADH-dependent MTAL effect is due to increased apical Na+/K+/2Cl- admittance and apical K+ recycling accompanied by a rise in calculated intracellular Cl- concentrations and by a threefold rise in basolateral Cl- conductance. rbClC-Ka, a 75.2 member of the ClC family of Cl- channels, mediates net Cl- absorption in the MTAL. The gating characteristics of rbClC-Ka channels from their intracellular surfaces are, to our knowledge, unique among Cl- channels. The channels are activated by small increases in intracellular Cl- (K( 1/2 ) = 10 mM Cl-). Adenosine triphosphate plus the catalytic subunit of protein kinase A (ATP + PKA) gate rbClC-Ka when cytosolic Cl- concentrations are 25 mM. Thus, in mouse MTAL segments, ADH-dependent rises in cytosolic Cl- are primarily responsible for basolateral Cl- conductance increases. Methods. These experiments compared the properties of Cl- channels fused into bilayers from basolaterally enriched vesicles from cultured mouse CTAL cells with rbClC-Ka channels. Results. The key findings were that anti-rbClC-Ka, an antibody that recognizes and blocks rbClC-Ka, recognized and blocked basolateral Cl- channels in CTAL cells, that the extracellular faces of the CTAL channels were, like rbClC-Ka, substrate gated with a K( 1/2 ) of approximately 170 mM Cl-, and that, unlike rbClC-Ka channels, cytosolic faces of basolateral CTAL Cl- channels were not gated by either increasing cytosolic Cl- concentrations or cytosolic (ATP + PKA). This failure of activation of basolateral CTAL Cl- channels was confirmed using excised patch clamp studies. Finally, on Western blots, anti-rbClC-Ka recognized a 74 kDa band on basolateral CTAL vesicles. Conclusions. Basolateral CTAL Cl- channels probably share a high degree of structural homology and possibly molecular mass with rbClC-Ka channels. However, significant differences between rbClC- Ka channels and CTAL Cl- channels account for the inability of increasing either cytosolic Cl- or (PKA + ATP) to raise P(o) in CTAL basolateral Cl- channels.

Original languageEnglish (US)
Pages (from-to)593-601
Number of pages9
JournalKidney international
Volume55
Issue number2
DOIs
StatePublished - 1999

Keywords

  • Chloride channels
  • Potassium recycling
  • Sodium chloride
  • Thick ascending limb
  • rbClC-Ka

ASJC Scopus subject areas

  • Nephrology

Fingerprint Dive into the research topics of 'Cl<sup>-</sup> channels in basolateral TAL membranes: XIII. Heterogeneity between basolateral MTAL and CTAL Cl<sup>-</sup> channels'. Together they form a unique fingerprint.

Cite this