Capacitance measurements of exocytosis in mouse pancreatic alpha-, beta- and delta-cells within intact islets of Langerhans

J Physiol. 2004 May 1;556(Pt 3):711-26. doi: 10.1113/jphysiol.2003.059675. Epub 2004 Feb 13.

Abstract

Capacitance measurements of exocytosis were applied to functionally identified alpha-, beta- and delta-cells in intact mouse pancreatic islets. The maximum rate of capacitance increase in beta-cells during a depolarization to 0 mV was equivalent to 14 granules s(-1), <5% of that observed in isolated beta-cells. Beta-cell secretion exhibited bell-shaped voltage dependence and peaked at +20 mV. At physiological membrane potentials (up to approximately -20 mV) the maximum rate of release was approximately 4 granules s(-1). Both exocytosis (measured by capacitance measurements) and insulin release (detected by radioimmunoassay) were strongly inhibited by the L-type Ca(2+) channel blocker nifedipine (25 microm) but only marginally (<20%) affected by the R-type Ca(2+) channel blocker SNX482 (100 nm). Exocytosis in the glucagon-producing alpha-cells peaked at +20 mV. The capacitance increases elicited by pulses to 0 mV exhibited biphasic kinetics and consisted of an initial transient (150 granules s(-1)) and a sustained late component (30 granules s(-1)). Whereas addition of the N-type Ca(2+) channel blocker omega-conotoxin GVIA (0.1 microm) inhibited glucagon secretion measured in the presence of 1 mm glucose to the same extent as an elevation of glucose to 20 mm, the L-type Ca(2+) channel blocker nifedipine (25 microm) had no effect. Thus, glucagon release during hyperglycaemic conditions depends principally on Ca(2+)-influx through N-type rather than L-type Ca(2+) channels. Exocytosis in the somatostatin-secreting delta-cells likewise exhibited two kinetically separable phases of capacitance increase and consisted of an early rapid (600 granules s(-1)) component followed by a sustained slower (60 granules s(-1)) component. We conclude that (1) capacitance measurements in intact pancreatic islets are feasible; (2) exocytosis measured in beta-cells in situ is significantly slower than that of isolated cells; and (3) the different types of islet cells exhibit distinct exocytotic features.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials / drug effects
  • Action Potentials / physiology
  • Animals
  • Calcium Channels, L-Type / drug effects
  • Calcium Channels, L-Type / physiology
  • Calcium Channels, N-Type / drug effects
  • Calcium Channels, N-Type / physiology
  • Calcium Channels, R-Type / drug effects
  • Calcium Channels, R-Type / physiology
  • Cells, Cultured
  • Electric Capacitance*
  • Electrophysiology
  • Exocytosis / drug effects
  • Exocytosis / physiology*
  • Glucagon / metabolism
  • Glucose / pharmacology
  • Insulin / metabolism
  • Insulin Secretion
  • Islets of Langerhans / cytology
  • Islets of Langerhans / drug effects
  • Islets of Langerhans / physiology*
  • Kinetics
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Mice
  • Mice, Inbred Strains
  • Microscopy, Electron, Transmission
  • Nifedipine / pharmacology
  • Patch-Clamp Techniques
  • Pertussis Toxin / pharmacology
  • Secretory Vesicles / ultrastructure
  • Somatostatin-Secreting Cells / cytology
  • Somatostatin-Secreting Cells / physiology*
  • Spider Venoms / pharmacology
  • omega-Conotoxin GVIA / pharmacology

Substances

  • Calcium Channels, L-Type
  • Calcium Channels, N-Type
  • Calcium Channels, R-Type
  • Insulin
  • SNX 482
  • Spider Venoms
  • Glucagon
  • omega-Conotoxin GVIA
  • Pertussis Toxin
  • Nifedipine
  • Glucose