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Cell Calcium. 1995 Feb;17(2):97-110.

Introduction of calcium buffers into the cytosol of Dictyostelium discoideum amoebae alters cell morphology and inhibits chemotaxis.

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1
Faculty of Biology, University of Konstanz, Germany.

Abstract

Differentiating Dictyostelium discoideum amoebae respond chemotactically towards the attractant cAMP. To test whether chemotaxis requires the establishment of a spatial gradient of the cytosolic calcium concentration ([Ca2+]i) we scrape-loaded calcium chelating agents with different affinities for Ca2+ into the cytosol of the cells. The buffers were 1,2-bis(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA) and its derivatives. Parameters analyzed were general cell morphology and the capability to protrude pseudopods and to migrate towards a cAMP-filled capillary. The chelators dose- and time-dependently inhibited spreading of the amoebae on the substrate. Both oriented pseudopod formation and locomotion of the cells were reduced. This effect was overcome by extracellular Ca2+, but not Mg2+. The effects of BAPTA derivatives were compared to the inhibition by BAPTA. A dose-response curve was obtained; 5,5'-difluoro-BAPTA was the most potent analogue. We conclude that a [Ca2+]i-gradient is necessary for orientation and locomotion. Chemotaxis experiments performed in the presence of extracellular EGTA revealed that liberation of Ca2+ from intracellular stores is sufficient for pseudopod formation; yet under physiological conditions influx of extracellular Ca2+ is also used to establish the gradient.

PMID:
7736566
[Indexed for MEDLINE]
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