Using light to shape chemical gradients for parallel and automated analysis of chemotaxis

Sean Collins, Hee Won Yang, Kimberly M. Bonger, Emmanuel G. Guignet, Thomas J. Wandless, Tobias Meyer

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Abstract Numerous molecular components have been identified that regulate the directed migration of eukaryotic cells toward sources of chemoattractant. However, how the components of this system are wired together to coordinate multiple aspects of the response, such as directionality, speed, and sensitivity to stimulus, remains poorly understood. Here we developed a method to shape chemoattractant gradients optically and analyze cellular chemotaxis responses of hundreds of living cells per well in 96-well format by measuring speed changes and directional accuracy. We then systematically characterized migration and chemotaxis phenotypes for 285 siRNA perturbations. A key finding was that the G-protein Giα subunit selectively controls the direction of migration while the receptor and Gβ subunit proportionally control both speed and direction. Furthermore, we demonstrate that neutrophils chemotax persistently in response to gradients of fMLF but only transiently in response to gradients of ATP. The method we introduce is applicable for diverse chemical cues and systematic perturbations, can be used to measure multiple cell migration and signaling parameters, and is compatible with low- and high-resolution fluorescence microscopy. Synopsis A new strategy, involving optical shaping of gradients, allows systematically analyzing components regulating cell migration speed and directionality. The approach is applied to characterize migration and chemotaxis phenotypes for 285 siRNA perturbations in human neutrophils. Automated uncaging of attractants allows systematic live-cell imaging of chemotaxis. Leukocytes have distinct components specialized for regulating cell speed and cell direction in response to chemoattractant gradients. Specialization in the chemoattractant signaling pathway occurs already at the level of the G-proteins. A new strategy, involving optical shaping of gradients, allows systematically analyzing components regulating cell migration speed and directionality. The approach is applied to characterize migration and chemotaxis phenotypes for 285 siRNA perturbations in human neutrophils.

Original languageEnglish (US)
Pages (from-to)1-13
Number of pages13
JournalMolecular Systems Biology
Volume11
Issue number4
DOIs
StatePublished - Apr 1 2015

Fingerprint

Chemotaxis
chemotaxis
chemoattractants
Chemotactic Factors
Migration
Gradient
Neutrophils
Cell Migration
Light
small interfering RNA
cell movement
Small Interfering RNA
Cell Movement
neutrophils
Phenotype
Cell
Perturbation
G Protein
G-proteins
phenotype

Keywords

  • chemokinesis
  • chemotaxis
  • Galphai
  • gradients
  • uncaging

ASJC Scopus subject areas

  • Information Systems
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Agricultural and Biological Sciences(all)
  • Computational Theory and Mathematics
  • Applied Mathematics

Cite this

Using light to shape chemical gradients for parallel and automated analysis of chemotaxis. / Collins, Sean; Yang, Hee Won; Bonger, Kimberly M.; Guignet, Emmanuel G.; Wandless, Thomas J.; Meyer, Tobias.

In: Molecular Systems Biology, Vol. 11, No. 4, 01.04.2015, p. 1-13.

Research output: Contribution to journalArticle

Collins, Sean ; Yang, Hee Won ; Bonger, Kimberly M. ; Guignet, Emmanuel G. ; Wandless, Thomas J. ; Meyer, Tobias. / Using light to shape chemical gradients for parallel and automated analysis of chemotaxis. In: Molecular Systems Biology. 2015 ; Vol. 11, No. 4. pp. 1-13.
@article{d12762cae7b845518a948ab2b9e048ba,
title = "Using light to shape chemical gradients for parallel and automated analysis of chemotaxis",
abstract = "Abstract Numerous molecular components have been identified that regulate the directed migration of eukaryotic cells toward sources of chemoattractant. However, how the components of this system are wired together to coordinate multiple aspects of the response, such as directionality, speed, and sensitivity to stimulus, remains poorly understood. Here we developed a method to shape chemoattractant gradients optically and analyze cellular chemotaxis responses of hundreds of living cells per well in 96-well format by measuring speed changes and directional accuracy. We then systematically characterized migration and chemotaxis phenotypes for 285 siRNA perturbations. A key finding was that the G-protein Giα subunit selectively controls the direction of migration while the receptor and Gβ subunit proportionally control both speed and direction. Furthermore, we demonstrate that neutrophils chemotax persistently in response to gradients of fMLF but only transiently in response to gradients of ATP. The method we introduce is applicable for diverse chemical cues and systematic perturbations, can be used to measure multiple cell migration and signaling parameters, and is compatible with low- and high-resolution fluorescence microscopy. Synopsis A new strategy, involving optical shaping of gradients, allows systematically analyzing components regulating cell migration speed and directionality. The approach is applied to characterize migration and chemotaxis phenotypes for 285 siRNA perturbations in human neutrophils. Automated uncaging of attractants allows systematic live-cell imaging of chemotaxis. Leukocytes have distinct components specialized for regulating cell speed and cell direction in response to chemoattractant gradients. Specialization in the chemoattractant signaling pathway occurs already at the level of the G-proteins. A new strategy, involving optical shaping of gradients, allows systematically analyzing components regulating cell migration speed and directionality. The approach is applied to characterize migration and chemotaxis phenotypes for 285 siRNA perturbations in human neutrophils.",
keywords = "chemokinesis, chemotaxis, Galphai, gradients, uncaging",
author = "Sean Collins and Yang, {Hee Won} and Bonger, {Kimberly M.} and Guignet, {Emmanuel G.} and Wandless, {Thomas J.} and Tobias Meyer",
year = "2015",
month = "4",
day = "1",
doi = "10.15252/msb.20156027",
language = "English (US)",
volume = "11",
pages = "1--13",
journal = "Molecular Systems Biology",
issn = "1744-4292",
publisher = "Nature Publishing Group",
number = "4",

}

TY - JOUR

T1 - Using light to shape chemical gradients for parallel and automated analysis of chemotaxis

AU - Collins, Sean

AU - Yang, Hee Won

AU - Bonger, Kimberly M.

AU - Guignet, Emmanuel G.

AU - Wandless, Thomas J.

AU - Meyer, Tobias

PY - 2015/4/1

Y1 - 2015/4/1

N2 - Abstract Numerous molecular components have been identified that regulate the directed migration of eukaryotic cells toward sources of chemoattractant. However, how the components of this system are wired together to coordinate multiple aspects of the response, such as directionality, speed, and sensitivity to stimulus, remains poorly understood. Here we developed a method to shape chemoattractant gradients optically and analyze cellular chemotaxis responses of hundreds of living cells per well in 96-well format by measuring speed changes and directional accuracy. We then systematically characterized migration and chemotaxis phenotypes for 285 siRNA perturbations. A key finding was that the G-protein Giα subunit selectively controls the direction of migration while the receptor and Gβ subunit proportionally control both speed and direction. Furthermore, we demonstrate that neutrophils chemotax persistently in response to gradients of fMLF but only transiently in response to gradients of ATP. The method we introduce is applicable for diverse chemical cues and systematic perturbations, can be used to measure multiple cell migration and signaling parameters, and is compatible with low- and high-resolution fluorescence microscopy. Synopsis A new strategy, involving optical shaping of gradients, allows systematically analyzing components regulating cell migration speed and directionality. The approach is applied to characterize migration and chemotaxis phenotypes for 285 siRNA perturbations in human neutrophils. Automated uncaging of attractants allows systematic live-cell imaging of chemotaxis. Leukocytes have distinct components specialized for regulating cell speed and cell direction in response to chemoattractant gradients. Specialization in the chemoattractant signaling pathway occurs already at the level of the G-proteins. A new strategy, involving optical shaping of gradients, allows systematically analyzing components regulating cell migration speed and directionality. The approach is applied to characterize migration and chemotaxis phenotypes for 285 siRNA perturbations in human neutrophils.

AB - Abstract Numerous molecular components have been identified that regulate the directed migration of eukaryotic cells toward sources of chemoattractant. However, how the components of this system are wired together to coordinate multiple aspects of the response, such as directionality, speed, and sensitivity to stimulus, remains poorly understood. Here we developed a method to shape chemoattractant gradients optically and analyze cellular chemotaxis responses of hundreds of living cells per well in 96-well format by measuring speed changes and directional accuracy. We then systematically characterized migration and chemotaxis phenotypes for 285 siRNA perturbations. A key finding was that the G-protein Giα subunit selectively controls the direction of migration while the receptor and Gβ subunit proportionally control both speed and direction. Furthermore, we demonstrate that neutrophils chemotax persistently in response to gradients of fMLF but only transiently in response to gradients of ATP. The method we introduce is applicable for diverse chemical cues and systematic perturbations, can be used to measure multiple cell migration and signaling parameters, and is compatible with low- and high-resolution fluorescence microscopy. Synopsis A new strategy, involving optical shaping of gradients, allows systematically analyzing components regulating cell migration speed and directionality. The approach is applied to characterize migration and chemotaxis phenotypes for 285 siRNA perturbations in human neutrophils. Automated uncaging of attractants allows systematic live-cell imaging of chemotaxis. Leukocytes have distinct components specialized for regulating cell speed and cell direction in response to chemoattractant gradients. Specialization in the chemoattractant signaling pathway occurs already at the level of the G-proteins. A new strategy, involving optical shaping of gradients, allows systematically analyzing components regulating cell migration speed and directionality. The approach is applied to characterize migration and chemotaxis phenotypes for 285 siRNA perturbations in human neutrophils.

KW - chemokinesis

KW - chemotaxis

KW - Galphai

KW - gradients

KW - uncaging

UR - http://www.scopus.com/inward/record.url?scp=84928788720&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84928788720&partnerID=8YFLogxK

U2 - 10.15252/msb.20156027

DO - 10.15252/msb.20156027

M3 - Article

C2 - 25908733

AN - SCOPUS:84928788720

VL - 11

SP - 1

EP - 13

JO - Molecular Systems Biology

JF - Molecular Systems Biology

SN - 1744-4292

IS - 4

ER -