Abstract
We have used the whole cell technique to microinject human fibroblasts with either 1,4,5-inositol trisphosphate (InsP 3) or ‘caged’ InsP 3 in order to study the mechanisms of transmembrane signalling related to mitogenic stimulations. Cytosolic Ca 2+ elevations in response to 1,4,5 InsP 3 diffusing from the patch pipette were difficult to detect, while 1,4,5 InsP 3, photoreleased after loading the cell with its inactive precursor, was capable of generating not only a single cytosolic Ca 2+ rise but sometimes triggered an oscillatory calcium response, similar to that often observed under mitogenic stimulation. We estimated that less than 100 nM InsP 3 was sufficient to generate Ca 2+ responses. The Ca 2+ rise produced by the photoreleased InsP 3 could fully activate the K + channels present in the plasma membrane of human fibroblasts.
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