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Ca(2+) signals in Pmr1-GFP-expressing COS-1 cells with functional endoplasmic reticulum
Authors:Missiaen Ludwig  Vanoevelen Jo  Van Acker Kristel  Raeymaekers Luc  Parys Jan B  Callewaert Geert  Wuytack Frank  De Smedt Humbert
Institution:Laboratorium voor Fysiologie, K.U. Leuven Campus Gasthuisberg, Herestraat 49, Leuven B-3000, Belgium. ludwig.missiaen@med.kuleuven.ac.be
Abstract:We studied the role of the Pmr1-containing Ca(2+) store in COS-1 cells endowed with a functional endoplasmic reticulum. Transfected cells could be recognized by using a green-fluorescent-protein (GFP)-tagged form of Pmr1. Pmr1-GFP fluorescence showed a typical juxtanuclear Golgi-like distribution. Pmr1-GFP-containing cells with functional endoplasmic reticulum responded to 100 microM ATP with baseline Ca(2+) spiking, while non-transfected cells produced an initial Ca(2+) peak followed by a long-lasting plateau. The Ca(2+) signal often appeared after a long latency in Pmr1-GFP-expressing cells. ATP-stimulated Pmr1-GFP-expressing cells with functional endoplasmic reticulum responded after a latency period to extracellular Ca(2+) with a regenerative Ca(2+) signal, while non-transfected control cells responded with an immediate slow rise in free cytosolic Ca(2+) concentration. These results demonstrate the importance of the Pmr1-containing Ca(2+) store in generating or modifying cellular Ca(2+) signals.
Keywords:Intracellular calcium  Calcium pump  Calcium store  Calcium oscillation
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