Analysis of directional cell migration on defined FN gradients: role of intracellular signaling molecules.
Directional cell migration on extracellular matrix (ECM) plays important roles in embryonic development and adult organisms. To study the mechanisms and signaling pathways involved in the regulation of directional cell migration, we created defined fibronectin (FN) gradients by using microfluidic systems. We found that fibroblasts exhibited haptotaxis towards higher FN concentration on the gradient. Furthermore, the net movements in the direction of FN gradients correlated with the increase in the slope of the gradient although the overall rate of migration was not correlated. Consistent with previous observations on the uniformly coated surface, local higher FN concentration led to reduced migration rate due to increased spreading. Upon transfection of N-WASP or activated Cdc42, but not FAK or Grb7, the cells showed increased directional migration. However, transfection of FAK, but not the other signaling molecules, led to an increase in the persistence of directional cell migration, which is dependent on the slope of the gradient as well as FAK interaction with PI3K. Together, these studies reveal some novel properties of directional cell migration on defined FN gradient and suggested a role for FAK signaling and N-WASP and Cdc42 in the differential regulation of the persistence and rate of directional cell migration.
Rhoads
Daniel S
DS
Department of Molecular Medicine, Cornell University, Ithaca, NY 14853, USA.
Guan
Jun-Lin
JL
D000818:Animals; D019898:Autocrine Communication / Q000187:drug effects; D016466:CHO Cells; D002465:Cell Movement / Q000187:drug effects*; D006224:Cricetinae; D003412:Cricetulus; D004058:Diffusion / Q000187:drug effects; D005353:Fibronectins / Q000378:metabolism / Q000494:pharmacology*; D051417:Focal Adhesion Kinase 1 / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects*; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal / Q000378:metabolism; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism
D005353:Fibronectins; D051317:Wiskott-Aldrich Syndrome Protein, Neuronal; D019869:Phosphatidylinositol 3-Kinases; D051417:Focal Adhesion Kinase 1; D020764:cdc42 GTP-Binding Protein