Characterization of the maitotoxin-activated cationic current from human skin fibroblasts

J Physiol. 2002 Jan 1;538(Pt 1):79-86. doi: 10.1113/jphysiol.2001.013036.

Abstract

The maitotoxin (MTX)-induced cationic current (I(mtx)) from human skin fibroblasts was characterized using the patch-clamp technique in whole-cell configuration. Under resting conditions (absence of MTX), the main current observed is produced by an outwardly rectifying K(+) channel which is inhibited by 1 mM TEA. The current reversal potential was -86 mV (n = 12). MTX (500 pM) activated a current with a linear current-voltage relationship and a reversal potential of -10 mV (n = 10). Replacing the extracellular Na(+) and K(+) with N-methyl-D-glucamine (NMDG) caused a shift of the reversal potential to a value below -100 mV, indicating that Na(+) and K(+), but not NMDG, carry I(mtx). Further ion selectivity experiments showed that Ca(2+) carries I(mtx) also. The resulting permeability sequence obtained with the Goldman-Hodgkin-Katz equation yielded Na(+) (1) approximately equal to K(+) (1) > Ca(2+) (0.87). The I(mtx) activation time course reflected the changes in intracellular Ca(2+) and Na(+) measured with the fluorescent indicators fura-2 and SBFI, respectively, suggesting that the activation of I(mtx) brings about an increment in intracellular Ca(2+) and Na(+). Reducing the extracellular Ca(2+) concentration below 1.8 mM prevented the activation of I(mtx) and the increment in intracellular Na(+) induced by MTX. Mn(2+) and Mg(2+) could not replace Ca(2+), but Ba(2+) could replace Ca(2+). MTX activation of current in 10 mM Ba(2+) was approximately 50 % of that induced in the presence of 1.8 mM Ca(2+). When 5 mM of the Ca(2+) chelator BAPTA was included in the patch pipette, MTX either failed to activate the current or induced a small current (less than 15 % of the control), indicating that intracellular Ca(2+) is also required for the activation of I(mtx). Intracellular Ba(2+) can replace Ca(2+) as an activator of I(mtx). However, in the presence of 10 mM Ba(2+) the activation by MTX of the current was 50 % less than the activation with nM concentrations of free intracellular Ca(2+).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Barium / physiology
  • Calcium / physiology
  • Cations, Divalent / metabolism*
  • Cells, Cultured
  • Electric Conductivity
  • Extracellular Space / metabolism
  • Fibroblasts / drug effects*
  • Fibroblasts / physiology*
  • Humans
  • Intracellular Membranes / metabolism
  • Ion Channels / physiology*
  • Magnesium / physiology
  • Marine Toxins / pharmacology*
  • Oxocins*
  • Patch-Clamp Techniques
  • Potassium / physiology
  • Skin / cytology
  • Skin Physiological Phenomena / drug effects*
  • Sodium / physiology

Substances

  • Cations, Divalent
  • Ion Channels
  • Marine Toxins
  • Oxocins
  • Barium
  • Sodium
  • maitotoxin
  • Magnesium
  • Potassium
  • Calcium