The contractile vacuole in Ca2+-regulation in Dictyostelium: its essential function for cAMP-induced Ca2+-influx
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Background: cAMP-induced Ca2+-influx in Dictyostelium is controlled by at least two non-mitochondrial Ca2+-stores: acidic stores and the endoplasmic reticulum (ER). The acidic stores may comprise the contractile vacuole network (CV), the endosomal compartment and acidocalcisomes. Here the role of CV in respect to function as a potential Ca2+-store was investigated. -- Results: Dajumin-GFP labeled contractile vacuoles were purified 7-fold by anti-GFP-antibodies in a magnetic field. The purified CV were shown for the first time to accumulate and release Ca2+. Release of Ca2+ was elicited by arachidonic acid or the calmodulin antagonist W7, the latter due to inhibition of the pump. The characteristics of Ca2+-transport and Ca2+-release of CV were compared to similarly purified vesicles of the ER labeled by calnexin-GFP. Since the CV proved to be a highly efficient Ca2+-compartment we wanted to know whether or not it takes part in cAMP-induced Ca2+-influx. We made use of the LvsA--mutant expected to display reduced Ca2+-transport due to loss of calmodulin. We found a severe reduction of cAMP-induced Ca2+-influx into whole cells. -- Conclusion: The contractile vacuoles in Dictyostelium represent a highly efficient acidic Ca2+-store that is required for cAMP-induced Ca2+-influx.
Dieter Malchow, Daniel F. Lusche, and Christian Schlatterer are with the Department of Biology, University of Konstanz, Konstanz, Germany, -- Arturo de Lozanne is with the Section of Molecular Cell Developmental Biology, University of Texas at Austin, Austin, Texas, USA, -- Annette Muller-Taubenberger is with the Max Planck Institute for Biochemistry, D-82152 Martinsried, Germany, and the Institute for Cell Biology (ABI), Ludwig Maximilians University Münche, München, Germany -- Daniel F. Lusche is with the WM Keck Research Facility, Department of Biological Sciences Iowa City, USA