title: Chemotactic cell trapping in controlled alternating gradient fields.
abstract: Directed cell migration toward spatio-temporally varying chemotactic stimuli requires rapid cytoskeletal reorganization. Numerous studies provide evidence that actin reorganization is controlled by intracellular redistribution of signaling molecules, such as the PI4,5P2/PI3,4,5P3 gradient. However, exploring underlying mechanisms is difficult and requires careful spatio-temporal control of external chemotactic stimuli. We designed a microfluidic setup to generate alternating chemotactic gradient fields for simultaneous multicell exposure, greatly facilitating statistical analysis. For a quantitative description of intracellular response dynamics, we apply alternating time sequences of spatially homogeneous concentration gradients across 300 μm, reorienting on timescales down to a few seconds. Dictyostelium discoideum amoebae respond to gradient switching rates below 0.02 Hz by readapting their migration direction. For faster switching, cellular repolarization ceases and is completely stalled at 0.1 Hz. In this "chemotactically trapped" cell state, external stimuli alternate faster than intracellular feedback is capable to respond by onset of directed migration. To investigate intracellular actin cortex rearrangement during gradient switching, we correlate migratory cell response with actin repolymerization dynamics, quantified by a fluorescence distribution moment of the GFP fusion protein LimEΔcc. We find two fundamentally different cell polarization types and we could reveal the role of PI3-Kinase for cellular repolarization. In the early aggregation phase, PI3-Kinase enhances the capability of D. discoideum cells to readjust their polarity in response to spatially alternating gradient fields, whereas in aggregation competent cells the effect of PI3-Kinase perturbation becomes less relevant.
authors.affiliation: Faculty of Physics and Center for NanoScience, Ludwig-Maximilians-Universität in Munich, Geschwister-Scholl-Platz 1, 80539 Munich, Germany.
authors.forename: Börn
authors.initials: B
authors.lastname: Meier
authors.forename: Alejandro
authors.initials: A
authors.lastname: Zielinski
authors.forename: Christoph
authors.initials: C
authors.lastname: Weber
authors.forename: Delphine
authors.initials: D
authors.lastname: Arcizet
authors.forename: Simon
authors.initials: S
authors.lastname: Youssef
authors.forename: Thomas
authors.initials: T
authors.lastname: Franosch
authors.forename: Joachim O
authors.initials: JO
authors.lastname: Rädler
authors.forename: Doris
authors.initials: D
authors.lastname: Heinrich
mesh_terms: D000199:Actins / Q000378:metabolism; D055592:Biophysical Phenomena; D002630:Chemotactic Factors / Q000008:administration & dosage; D002633:Chemotaxis / Q000187:drug effects / Q000502:physiology*; D003470:Culture Media; D004023:Dictyostelium / Q000187:drug effects / Q000502:physiology; D020342:Finite Element Analysis; D046210:Microfluidic Analytical Techniques; D008954:Models, Biological; D009068:Movement / Q000502:physiology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D015800:Protozoan Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
chemical_list: D000199:Actins; D002630:Chemotactic Factors; D003470:Culture Media; D015800:Protozoan Proteins; D019869:Phosphatidylinositol 3-Kinases