Intracellular uptake and α-amylase and lactate dehydrogenase releasing actions of the divalent cation ionophore A23187 in dissociated pancreatic acinar cells

Douglas E. Chandler, John A. Williams

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

Intracellular uptake of A23187 and the increased release of amylase and lactate dehydrogenase (LDH) accompanying ionophore uptake was studied using dissociated acinar cells prepared from mouse pancreas. Easily detected changes in the fluorescence excitation spectrum of A23187 upon transfer of the ionophore from a Tris-buffered Ringer's to cell membranes were used to monitor A23187 uptake. Uptake was rapid in the absence of extracellular Ca2+ and Mg2+ (t1/2=1 min) and much slower in the presence of Ca2+ or Mg2+ (t1/2=20 min). Cell-associated ionophore was largely intracellular as indicated by fluorescence microscopy, lack of spectral sensitivity to changes in extracellular Ca2+ and Mg2+, and by equivalent interaction of ionophore with membranes of whole and sonicated cells. A23187 (10 μm) increased amylase release 200% in the presence of extracellular Ca2+ and Mg2+. In the absence of Ca2+ (but in the presence of Mg2+) A23187 did not increase amylase release. A23187 (10 μm) also produced Ca2+-dependent cell damage, as judged by increased LDH release, increased permeability to trypan blue, and by disruption of cell morphology. The cell damaging and amylase releasing properties of A23187 were distinguished by their time course and dose-response relationship. A23187 (1 μm) increased amylase release 140% without increasing LDH release or permeability to trypan blue.

Original languageEnglish (US)
Pages (from-to)201-230
Number of pages30
JournalThe Journal of Membrane Biology
Volume32
Issue number1
DOIs
StatePublished - Dec 1977
Externally publishedYes

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Acinar Cells
Ionophores
Divalent Cations
Calcimycin
Amylases
L-Lactate Dehydrogenase
Trypan Blue
Permeability
Fluorescence Microscopy
Pancreas
Fluorescence
Cell Membrane
Membranes

ASJC Scopus subject areas

  • Physiology
  • Cell Biology
  • Biophysics

Cite this

Intracellular uptake and α-amylase and lactate dehydrogenase releasing actions of the divalent cation ionophore A23187 in dissociated pancreatic acinar cells. / Chandler, Douglas E.; Williams, John A.

In: The Journal of Membrane Biology, Vol. 32, No. 1, 12.1977, p. 201-230.

Research output: Contribution to journalArticle

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