Genetic deletion of the Rho GEF Net1 impairs mouse macrophage motility and actin cytoskeletal organization

Small GTPases. 2020 Jul;11(4):293-300. doi: 10.1080/21541248.2017.1405772. Epub 2017 Dec 31.


Macrophages are innate immune cells that constantly patrol an organism to fulfill protective and homeostatic roles. Previous studies have shown that Rho GTPase activity is required for macrophage mobility, yet the roles of upstream regulatory proteins controlling Rho GTPase function in these cells are not well defined. Previously we have shown that the RhoA GEF Net1 is required for human breast cancer cell motility and extracellular matrix invasion. To assess the role of Net1 in macrophage motility, we isolated bone marrow macrophage (BMM) precursors from wild type and Net1 knockout mice. Loss of Net1 did not affect the ability of BMM precursors to differentiate into mature macrophages in vitro, as measured by CD68 and F4/80 staining. However, Net1 deletion significantly reduced RhoA activation, F-actin accumulation, adhesion, and motility in these cells. Nevertheless, similar to RhoA/RhoB double knockout macrophages, Net1 deletion did not impair macrophage recruitment to the peritoneum in a mouse model of sterile inflammation. These data demonstrate that Net1 is an important regulator of RhoA signaling and motility in mouse macrophages in vitro, but that its function may be dispensable for macrophage recruitment to inflammatory sites in vivo.

Keywords: Net1; RhoA; actin; adhesion; macrophage; motility.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Actins / metabolism*
  • Animals
  • Cell Differentiation
  • Cells, Cultured
  • Cytoskeleton / metabolism*
  • Gene Deletion
  • Macrophages / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Oncogene Proteins / deficiency
  • Oncogene Proteins / genetics*
  • Oncogene Proteins / metabolism
  • Rho Guanine Nucleotide Exchange Factors / genetics*
  • Rho Guanine Nucleotide Exchange Factors / metabolism


  • Actins
  • Net1 protein, mouse
  • Oncogene Proteins
  • Rho Guanine Nucleotide Exchange Factors