Shear stress-induced volume decrease in C11-MDCK cells by BK-α/β4

J. David Holtzclaw, Liping Liu, P. Richard Grimm, Steven C. Sansom

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

Large-conductance, calcium-activated potassium channels (BK) are expressed in principal cells (PC) and intercalated cells (IC) in mammalian nephrons as BK-α/β1 and BK-α/β4, respectively. IC, which protrude into the lumens of tubules, express substantially more BK than PC despite lacking sufficient Na-K-ATPase to support K secretion. We previously showed in mice that IC exhibit size reduction when experiencing high distal flows induced by a high-K diet. We therefore tested the hypothesis that BK-α/β4 are regulators of IC volume via a shear stress (τ)-induced, calcium-dependent mechanism, resulting in a reduction in intracellular K content. We determined by Western blot and immunocytochemical analysis that C11-Madin-Darby canine kidney cells contained a predominance of BK-α/β4. To determine the role of BK-α/β4 in τ-induced volume reduction, we exposed C11 cells to τ and measured K efflux by flame photometry and cell volume by calcein staining, which changes inversely to cell volume. With 10 dynes/cm2, calcein intensity significantly increased 39% and monovalent cationic content decreased significantly by 37% compared with static conditions. Furthermore, the shear-induced K loss from C11 was abolished by the reduction of extracellular calcium, addition of 5 mM TEA, or BK-β4 small interfering (si) RNA, but not by addition of nontarget siRNA. These results show that BK-α/β4 plays a role in shear-induced K loss from IC, suggesting that BK-α/β4 regulate IC volume during high-flow conditions. Furthermore, these results support the use of C11 cells as in vitro models for studying BK-related functions in IC of the kidney.

Original languageEnglish (US)
Pages (from-to)F507-F516
JournalAmerican Journal of Physiology - Renal Physiology
Volume299
Issue number3
DOIs
StatePublished - Sep 2010

Keywords

  • Calcein
  • Intercalated cells
  • Mechanotransduction
  • Parallel plate flow chamber
  • Potassium
  • Volume regulation

ASJC Scopus subject areas

  • Physiology
  • Urology

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