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A Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable Concentration Gradients
09:28

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Published on: April 19, 2010

Gradient sensing in defined chemotactic fields.

Monica Skoge1, Micha Adler, Alex Groisman

  • 1Division of Biological Sciences, University of California, La Jolla, CA 92093, USA.

Integrative Biology : Quantitative Biosciences From Nano to Macro
|October 1, 2010
PubMed
Summary
This summary is machine-generated.

Dictyostelium discoideum cells exhibit exquisite chemotaxis to cyclic adenosine monophosphate (cAMP) gradients. This study reveals how activated Ras (RasGTP) localization at pseudopod tips drives directional cell movement in response to cAMP signaling.

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Imaging G-protein Coupled Receptor (GPCR)-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum
09:40

Imaging G-protein Coupled Receptor (GPCR)-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum

Published on: September 20, 2011

Area of Science:

  • Cell Biology
  • Biochemistry
  • Microfluidics

Background:

  • Cells respond to external molecular cues, altering physiology and movement.
  • Motile cells like Dictyostelium discoideum use chemotaxis to navigate chemical gradients.
  • The mechanisms behind Dictyostelium's high sensitivity to cyclic adenosine monophosphate (cAMP) gradients remain largely unknown.

Purpose of the Study:

  • To investigate the chemotactic response of Dictyostelium discoideum cells to stable linear cAMP gradients.
  • To elucidate the role of activated Ras (RasGTP) in sensing and responding to cAMP gradients.

Main Methods:

  • Utilized microfluidic devices with low-ceiling gradient chambers to generate precise cAMP gradients.
  • Employed confocal microscopy to track the subcellular localization of RasGTP using a Ras Binding Domain fused to Green Fluorescent Protein (RBD-GFP).
  • Observed cell behavior and protein localization in response to varying cAMP gradient steepness and cessation.

Main Results:

  • Dictyostelium cells adapted to low-ceiling chambers by flattening and elongating, maintaining rapid movement.
  • Activated Ras (RasGTP) rapidly localized to the tips of extending pseudopods in response to cAMP.
  • RasGTP patches disassembled quickly when pseudopods stalled or the cAMP gradient was removed, indicating dynamic signaling.

Conclusions:

  • RasGTP localization dynamics correlate with pseudopod extension and cell movement direction.
  • These findings provide insights into signal transduction mechanisms converting weak external cAMP signals into strong directional bias.
  • Understanding RasGTP dynamics is crucial for deciphering the exquisite sensitivity of Dictyostelium chemotaxis.