Pb2(+)-induced secretion from bovine chromaffin cells: fura-2 as a probe for Pb2+

Am J Physiol. 1990 Nov;259(5 Pt 1):C762-8. doi: 10.1152/ajpcell.1990.259.5.C762.

Abstract

The effect of Pb2+ on catecholamine release was studied in isolated intact and permeabilized bovine chromaffin cells. Fura-2 was used to monitor intracellular Pb2+. A characterization of Pb2(+)-fura-2 interactions in solutions simulating intracellular ionic composition showed that Pb2+ forms a 1:1 Pb2(+)-fura-2 complex (apparent dissociation constant = 4.2 x 10(-12) M, pH 7.05) whose fluorescence resembles that of the Ca2(+)-fura-2 complex. Spectra recorded from fura-2-loaded cells indicate entry of Pb2+ into the cells. Intracellular Pb2+ concentrations were proportional to extracellular Pb2+ concentrations and were found to be 10(-11)-10(-12) M in cells exposed to micromolar Pb2+ concentrations. Pb2+ elicited the release of tritiated norepinephrine from fura-2-loaded cells, indicating the extraordinary effectiveness of Pb2+ as a releasing agent. Permeabilization of cells with digitonin showed that Pb2+ is able, in the absence of Ca2+, to produce exocytotic release at concentrations of 3.2 x 10(-10) M or higher (3 orders of magnitude lower than Ca2+). These results support the notion that Pb2+ can act as a potent Ca2+ surrogate in triggering secretion.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adrenal Medulla / drug effects
  • Adrenal Medulla / metabolism*
  • Animals
  • Calcium / pharmacology
  • Cattle
  • Cell Membrane Permeability
  • Fura-2
  • In Vitro Techniques
  • Kinetics
  • Lead / pharmacology*
  • Mathematics
  • Models, Theoretical
  • Norepinephrine / metabolism*
  • Spectrometry, Fluorescence

Substances

  • Lead
  • Calcium
  • Fura-2
  • Norepinephrine