Biology subjects

Paulson, S. G.

Publications and source records attributed to Paulson, S. G..

3 recordsLinked to original sources

bioRxiv2025

The Arp2/3 complex is required for in situ haptotactic response of microglia to iC3b

Microglia maintain brain homeostasis by performing iC3b-mediated synaptic pruning on excessive dendritic spines during neurodevelopment. Cellular interaction with iC3b is mediated by the Arp2/3 complex in other cell types, but understudied in microglia. Using a combination of in vitro and in situ physical confinement studies, we examined CR3-dependent clearance of iC3b in microglia and the role the Arp2/3 complex plays in enabling this clearance. We demonstrated Arp2/3 complex inhibition decreased phagocytosis and cell motility in vitro. Furthermore, we demonstrate that microglia-like cells are able to remove immobilized iC3b from the substrate in an Arp2/3-dependent fashion, in a process reminiscent of trogocytic synaptic pruning. We also used a novel approach to immobilize an iC3b gradient onto a substrate and demonstrate Arp2/3-dependent haptotactic migration toward increasing iC3b concentrations. Microglia demonstrate a persistent inability to stably interact with iC3b-coated beads in hippocampal slice cultures upon Arp2/3 complex inhibition. As a whole, the present study establishes new approaches to systematically interrogate molecular pathways relevant to synaptic pruning, advances the understanding of iC3b phagocytosis as a haptotactic response, and confirms that the Arp2/3-dependent haptotactic response is relevant for microglia in their normal physiological microenvironment.

neuroscience↗

bioRxiv2025

Macrophages migrate persistently and directionally upon entering 2D confinement in the presence of extracellular matrix

Cells sense and respond to their environment in a myriad of ways. In many instances they must integrate simultaneous cues ranging from the physical properties and composition of the extracellular matrix to guidance cues that stimulate chemotaxis or haptotaxis. How cells make sense of multiple simultaneous cues is an ongoing physiologically relevant question. The present study seeks to contribute to the understanding of multi-cue sensing by understanding how the transition to a confined setting with or without an added haptotactic gradient alters macrophage migration. We found that the transition to confinement is itself a directional cue capable of driving persistent migration hours after macrophages enter the confined environment. Next, we found that a haptotactic fibronectin gradient made cells even more directionally persistent under confinement. Finally, Arp2/3 complex deletion rendered macrophages unresponsive to the haptotactic gradient, but they retained directionally persistent migration due to their transition to confinement. These findings may be particularly relevant for cells that move from an adherent 2D environment into a confining 3D environment, like leukocytes and circulating tumor cells that extravasate into peripheral tissue. Summary StatementMacrophages migrate persistently after they transition from 2D adhesion to an adhesive confined environment. Migration is enhanced further if cells sense a fibronectin gradient. This may have relevance to immune and cancer cell behavior.

cell biology↗

bioRxiv2025

Physical confinement and phagocytic uptake induce directional migration

Physical confinement is not routinely considered as a factor that influences phagocytosis, which is typically assayed in unconfined settings in vitro. BV-2 microglia-like cells were used to interrogate the impact of confinement on IgG-mediated phagocytosis side by side with unconfined cells. Confinement acted as a potent phagocytic driver, greatly increasing the fraction of phagocytic cells in the population compared to the unconfined setting. Arp2/3 complex and myosin II contributed to this effect. Remarkably, confinement partially rescued phagocytic uptake upon myosin II disruption. In addition, cells under confinement were partially resistant to the actin-depolymerizing drug cytochalasin D. Unexpectedly, we observed that bead uptake stimulated persistent migration, a process we term phagocytic priming. Integrin-dependent adhesion was required for phagocytic priming in unconfined and confined settings, but was dispensable for phagocytic uptake. The cytoskeletal requirements for phagocytic priming differed depending on confinement state. Myosin II and Arp2/3 complex were required for phagocytic priming under confinement, but not in unconfined settings. As with phagocytosis, cytoskeleton-dependent priming of motility shifts depending on physical confinement status. Phagocytic priming may facilitate innate immune function by driving cells to more efficiently surveil their local microenvironment in response to wounds or trauma.

cell biology↗