Properties of voltage-gated Ca2+ currents measured from mouse pancreatic β-cells in situ

• DAVID MEARS ^[1] ; EDUARDO ROJAS ^[1]
  1. [1] Universidad de Chile

     Universidad de Chile

     Santiago, Chile

     
• Localización: Biological Research, ISSN-e 0717-6287, ISSN 0716-9760, Vol. 39, Nº. 3, 2006, págs. 505-520
• Idioma: inglés
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• Resumen

  • We used the single-microelectrode voltage-clamp technique to record ionic currents from pancreatic β-cells within intact mouse islets of Langerhans at 37C, the typical preparation for studies of glucose-induced "bursting" electrical activity. Cells were impaled with intracellular microelectrodes, and voltage pulses were applied in the presence of tetraethylammonium. Under these conditions, a voltage-dependent Ca2+ current (I Cav), containing L-type and non-L-type components, was observed. The current measured in situ was larger than that measured in single cells with whole-cell patch clamping, particularly at membrane potentials corresponding to the action potentials of β-cell electrical activity. The temperature dependence of I Cav was not sufficient to account for the difference in size of the currents recorded with the two methods. During prolonged pulses, the voltage-dependent Ca2+ current measured in situ displayed both rapid and slow components of inactivation. The rapid component was Ca2+-dependent and was inhibited by the membrane-permeable Ca2+ chelator, BAPTA-AM. The effect of BAPTA-AM on β-cell electrical activity then demonstrated that Ca2+-dependent inactivation of I Cav contributes to action potential repolarization and to control of burst frequency. Our results demonstrate the utility of voltage clamping β-cells in situ for determining the roles of ion channels in electrical activity and insulin secretion.