Biomedical Microdevices

, 17:100 | Cite as

Integrin associated proteins differentially regulate neutrophil polarity and directed migration in 2D and 3D

  • Yukie Yamahashi
  • Peter J. Cavnar
  • Laurel E. Hind
  • Erwin Berthier
  • David A. Bennin
  • David Beebe
  • Anna Huttenlocher


Directed neutrophil migration in blood vessels and tissues is critical for proper immune function; however, the mechanisms that regulate three-dimensional neutrophil chemotaxis remain unclear. It has been shown that integrins are dispensable for interstitial three-dimensional (3D) leukocyte migration; however, the role of integrin regulatory proteins during directed neutrophil migration is not known. Using a novel microfluidic gradient generator amenable to 2D and 3D analysis, we found that the integrin regulatory proteins Kindlin-3, RIAM, and talin-1 differentially regulate neutrophil polarization and directed migration to gradients of chemoattractant in 2D versus 3D. Both talin-1-deficient and RIAM-deficient neutrophil-like cells had impaired adhesion, polarization, and migration on 2D surfaces whereas in 3D the cells polarized but had impaired 3D chemotactic velocity. Kindlin-3 deficient cells were able to polarize and migrate on 2D surfaces but had impaired directionality. In a 3D environment, Kindlin-3 deficient cells displayed efficient chemotaxis. These findings demonstrate that the role of integrin regulatory proteins in cell polarity and directed migration can be different in 2D and 3D.


Neutrophil Chemotaxis Integrin regulatory proteins Microfluidics 



This work was supported by National Institutes of Health Grant RO1EB010039 (to A. H. and D. J. B.), AHA Grant 10POST3230031 (to P. J. C.), and University of Wisconsin Carbone Cancer Center Cancer Center, Support Grant P30 CA014520 (to D.J.B).


David J. Beebe holds equity in Bellbrook Labs, LLC, Tasso, Inc., and Salus Discovery, LLC. E. Berthier holds equity in Salus Discovery, LLC and Tasso, Inc. P. Cavnar holds equity in Salus Discovery, LLC.

Author contributions

YY, PC, EB, DB and AH designed the studies, analyzed data, and wrote the manuscript. YY, DAB, PC, EB performed experiments and analyzed data.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Yukie Yamahashi
    • 1
    • 2
  • Peter J. Cavnar
    • 3
  • Laurel E. Hind
    • 2
  • Erwin Berthier
    • 4
  • David A. Bennin
    • 1
    • 2
  • David Beebe
    • 4
  • Anna Huttenlocher
    • 1
    • 2
  1. 1.Department of PediatricsUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Department of Medical Microbiology and ImmunologyUniversity of Wisconsin-MadisonMadisonUSA
  3. 3.Department of BiologyUniversity of West FloridaPensacolaUSA
  4. 4.Department of BioengineeringUniversity of Wisconsin-MadisonMadisonUSA

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