Integrin associated proteins differentially regulate neutrophil polarity and directed migration in 2D and 3D.
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.
Department of Pediatrics, University of Wisconsin-Madison, Madison, WI, USA.|Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, 1550 Linden Drive, Room 4205 MSB, Madison, WI, 53706, USA.
Yukie
Y
Yamahashi
Department of Biology, University of West Florida, Pensacola, FL, USA.
Peter J
PJ
Cavnar
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, 1550 Linden Drive, Room 4205 MSB, Madison, WI, 53706, USA.
Laurel E
LE
Hind
Department of Bioengineering, University of Wisconsin-Madison, Madison, WI, USA.
Erwin
E
Berthier
Department of Pediatrics, University of Wisconsin-Madison, Madison, WI, USA.|Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, 1550 Linden Drive, Room 4205 MSB, Madison, WI, 53706, USA.
David A
DA
Bennin
Department of Bioengineering, University of Wisconsin-Madison, Madison, WI, USA.
David
D
Beebe
Department of Pediatrics, University of Wisconsin-Madison, Madison, WI, USA. huttenlocher@wisc.edu.|Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, 1550 Linden Drive, Room 4205 MSB, Madison, WI, 53706, USA. huttenlocher@wisc.edu.
Anna
A
Huttenlocher
D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D016764:Cell Polarity / Q000502:physiology; D002469:Cell Separation / Q000295:instrumentation; D002633:Chemotaxis / Q000502:physiology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017022:Flow Injection Analysis / Q000295:instrumentation*; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D056656:Lab-On-A-Chip Devices; D008904:Miniaturization; D009504:Neutrophils / Q000166:cytology* / Q000502:physiology*
D016023:Integrins
Chemotaxis; Integrin regulatory proteins; Microfluidics; Neutrophil