Genetic deletion of the Rho GEF <i>Net1</i> impairs mouse macrophage motility and actin cytoskeletal organization

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 <i>Net1</i> knockout mice. Loss of <i>Net1</i> did not affect the ability of BMM precursors to differentiate into mature macrophages <i>in vitro</i>, as measured by CD68 and F4/80 staining. However, <i>Net1</i> deletion significantly reduced RhoA activation, F-actin accumulation, adhesion, and motility in these cells. Nevertheless, similar to <i>RhoA/RhoB</i> double knockout macrophages, <i>Net1</i> 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 <i>in vitro</i>, but that its function may be dispensable for macrophage recruitment to inflammatory sites <i>in vivo</i>.