On identifying information from image-based spatial polarity phenotypes in neutrophils

Cell polarity is involved in many biological functions such as development, wound healing and immune responses. In human neutrophils, polarization is characterized by the translocation of distinct sets of signaling molecules to opposite ends of the cell and the rapid rearrangement of cytoskeleton to initiate migration. While many image-based studies have described cellular morphology and the intensity level of polarity signaling molecules, systematic characterization of the spatial distribution of polarity signaling molecules has been lacking. Here we designed a collection of analytical features to quantify spatial phenotypes of polarity molecules. We compared our features to commonly used polarity readouts and found that they captured additional aspects of the polarization dynamics that were not contained in the existing features. Our work provides a starting point to identify informative features for the study of neutrophil polarization.

[1]  C. Bakal,et al.  Quantitative Morphological Signatures Define Local Signaling Networks Regulating Cell Morphology , 2007, Science.

[2]  W. Raub.  From the National Institutes of Health. , 1990, JAMA.

[3]  Lani F. Wu,et al.  Image-based multivariate profiling of drug responses from single cells , 2007, Nature Methods.

[4]  S. Gygi,et al.  Hem-1 Complexes Are Essential for Rac Activation, Actin Polymerization, and Myosin Regulation during Neutrophil Chemotaxis , 2006, PLoS biology.

[5]  G. Danuser,et al.  Two Distinct Actin Networks Drive the Protrusion of Migrating Cells , 2004, Science.

[6]  Olivier Pertz,et al.  Neutrophil polarization: spatiotemporal dynamics of RhoA activity support a self-organizing mechanism. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Jingsong Xu,et al.  Divergent Signals and Cytoskeletal Assemblies Regulate Self-Organizing Polarity in Neutrophils , 2003, Cell.

[8]  B. Helmke,et al.  Spatiotemporal Analysis of Actin Ruffling Dynamics in Living Cells , 2006, 2006 Fortieth Asilomar Conference on Signals, Systems and Computers.

[9]  Natalie Andrew,et al.  Chemotaxis in shallow gradients is mediated independently of PtdIns 3-kinase by biased choices between random protrusions , 2007, Nature Cell Biology.

[10]  Mark M Davis,et al.  Quantitative imaging of lymphocyte membrane protein reorganization and signaling. , 2005, Biophysical journal.

[11]  Richard A. Firtel,et al.  G protein–independent Ras/PI3K/F-actin circuit regulates basic cell motility , 2007, The Journal of cell biology.

[12]  T. Meyer,et al.  A local coupling model and compass parameter for eukaryotic chemotaxis. , 2005, Developmental cell.

[13]  A. Bøyum,et al.  Isolation of mononuclear cells and granulocytes from human blood. , 1968 .

[14]  M V Boland,et al.  After sequencing: quantitative analysis of protein localization. , 1999, IEEE engineering in medicine and biology magazine : the quarterly magazine of the Engineering in Medicine & Biology Society.

[15]  R. Murphy,et al.  Engineering in genomics , 1999, IEEE Engineering in Medicine and Biology Magazine.