Synchronization of Dictyostelium discoideum adhesion and spreading using electrostatic forces

Synchronization of cell spreading is valuable for the study of molecular events involved in the formation of adhesive contacts with the substrate. At a low ionic concentration (0.17 mM) Dictyostelium discoideum cells levitate over negatively charged surfaces due to electrostatic repulsion. First, a...

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Published in:Bioelectrochemistry (Amsterdam, Netherlands) Netherlands), 2010-10, Vol.79 (2), p.198-210
Main Authors: Socol, Marius, Lefrou, Christine, Bruckert, Franz, Delabouglise, Didier, Weidenhaupt, Marianne
Format: Article
Language:English
Subjects:
Actins - metabolism
Biotechnology
Cell adhesion
Cell Adhesion - physiology
Cell Movement - physiology
Chemical Sciences
Computer Science
Dictyostelium - cytology
Dictyostelium - physiology
Dictyostelium discoideum
Electrochemistry
Electrophysiological Phenomena
Electrostatic forces
Fluorescence
Green Fluorescent Proteins - analysis
Indium Tin Oxide
Intracellular Membranes - metabolism
Life Sciences
Lim EΔcoil fluorescence
Material chemistry
Microscopy, Interference
Reflection Interference Contrast Microscopy
Static Electricity
Tin Compounds - chemistry
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Summary:Synchronization of cell spreading is valuable for the study of molecular events involved in the formation of adhesive contacts with the substrate. At a low ionic concentration (0.17 mM) Dictyostelium discoideum cells levitate over negatively charged surfaces due to electrostatic repulsion. First, a two-chamber device, divided by a porous membrane, allows to quickly increase the ionic concentration around the levitating cells. In this way, a good synchronization was obtained, the onsets of cell spreading being separated by less than 5 s. Secondly applying a high potential pulse (2.5 V/Ref, 0.1 s) to an Indium Tin Oxide surface attracts the cells toward the surface where they synchronously spread as monitored by Lim E Δcoil-GFP. During spreading, actin polymerizes in series of active spots. On average, the first spot appears 8–11 s after the electric pulse and the next ones appear regularly, separated by about 10 s. Synchronized actin-polymerization activity continues for 40 s. Using an electric pulse to control the exact time point at which cells contact the surface has allowed for the first time to quantify the cellular response time for actin polymerization. Electrochemical synchronization is therefore a valuable tool to study intracellular responses to contact.
ISSN:1567-5394
1878-562X
DOI:10.1016/j.bioelechem.2010.04.003