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J Biol Chem, Vol. 275, Issue 18, 13411-13414, May 5, 2000

Ca²⁺ Depletion and Inositol 1,4,5-Trisphosphate-evoked Activation of Ca²⁺ Entry in Single Guinea Pig Hepatocytes^*

Gilles Guihard, Jacques Noel^§, and Thierry Capiod^¶

From INSERM U442, Université Paris-Sud, Bât. 443, 91405 Orsay, France

Store-operated Ca²⁺ entry was investigated by monitoring the Ca²⁺-dependent K⁺ permeability in voltage-clamped guinea pig hepatocytes. In physiological conditions, intracellular Ca²⁺ stores are discharged following agonist stimulation, but depletion of this stores can be achieved using Ca²⁺-Mg²⁺-ATPase inhibitors such as 2,5-di(tert-butyl)-1,4-benzohydroquinone and thapsigargin. The effect of internal Ca²⁺ store depletion on Ca²⁺ influx was tested in single cells using inositol 1,4,5-trisphosphate (InsP₃) release from caged InsP₃ after treatment of the cells with 2,5-di(tert-butyl)-1,4-benzohydroquinone or thapsigargin in Ca²⁺-free solutions. We show that the photolytic release of 1-D-myo-inositol 1,4-bisphosphate 5-phosphorothioate, a stable analog of InsP₃, and Ca²⁺ store depletion have additive effects to activate a high level of Ca²⁺ entry in single guinea pig hepatocytes. These results suggest that there is a direct functional interaction between InsP₃ receptors and Ca²⁺ channels in the plasma membrane, although the nature of these Ca²⁺ channels in hepatocytes is unclear.

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