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Microglia, the resident immune cells of the brain, are crucial in the development of the nervous system. Recent evidence demonstrates that microglia modulate adult hippocampal neurogenesis by inhibiting cell proliferation of neural precursors and survival both in vitro and in vivo, thus maintaining a balance between cell division and cell death in the neural stem cell pool. There are increasing reports suggesting these microglia found in neurogenic niches differ from their counterparts in non-neurogenic areas. Here, we present evidence that microglia in the hippocampal neurogenic niche are a specialized population that express genes known to regulate neurogenesis. By comprehensively profiling myeloid lineage cells in the hippocampus using single cell RNA-sequencing, we resolve transcriptomic differences in microglia originating from the subgranular zone. These cells have lower expression of genes associated with homeostatic microglia and increased expression of genes associated with phagocytosis. Intriguingly, this small yet distinct population expresses a gene signature with substantial overlap with previously characterized phenotypes, including disease associated microglia (DAM), a particularly unique and compelling microglial state.

For efficient polarity and migration, cells need to regulate the magnitude and spatial distribution of actin assembly. This process is coordinated by reciprocal interactions between the actin cytoskeleton and mechanical forces. Actin polymerization-based protrusion increases tension in the plasma membrane, which in turn acts as a long-range inhibitor of actin assembly. These interactions form a negative feedback circuit that limits the magnitude of membrane tension in neutrophils and prevents expansion of the existing front and the formation of secondary fronts. It has been suggested that the plasma membrane directly inhibits actin assembly by serving as a physical barrier that opposes protrusion. Here we show that efficient control of actin polymerization-based protrusion requires an additional mechanosensory feedback cascade that indirectly links membrane tension with actin assembly. Specifically, elevated membrane tension acts through phospholipase D2 (PLD2) and the mammalian target of rapamycin complex 2 (mTORC2) to limit actin nucleation. In the absence of this pathway, neutrophils exhibit larger leading edges, higher membrane tension, and profoundly defective chemotaxis. Mathematical modeling suggests roles for both the direct (mechanical) and indirect (biochemical via PLD2 and mTORC2) feedback loops in organizing cell polarity and motility—the indirect loop is better suited to enable competition between fronts, whereas the direct loop helps spatially organize actin nucleation for efficient leading edge formation and cell movement. This circuit is essential for polarity, motility, and the control of membrane tension., A mechanosensory biochemical cascade involving phospholipase D2 and mTORC2 coordinates physical forces and cytoskeletal rearrangements to allow efficient polarization and migration of neutrophils., Author Summary How cells regulate the size and number of their protrusions for efficient polarity and motility is a fundamental question in cell biology. We recently found that immune cells known as neutrophils use physical forces to regulate this process. Actin polymerization-based protrusion stretches the plasma membrane, and this increased membrane tension acts as a long-range inhibitor of actin-based protrusions elsewhere in the cell. Here we investigate how membrane tension limits protrusion. We demonstrate that the magnitude of actin network assembly in neutrophils is determined by a mechanosensory biochemical cascade that converts increases in membrane tension into decreases in protrusion. Specifically, we show that increasing plasma membrane tension acts through a pathway containing the phospholipase D2 (PLD2) and the mammalian target of rapamycin complex 2 (mTORC2) to limit actin network assembly. Without this negative feedback pathway, neutrophils exhibit larger leading edges, higher membrane tension, and profoundly defective chemotaxis. Mathematical modeling indicates that this feedback circuit is a favorable topology to enable competition between protrusions during neutrophil polarization. Our work shows how biochemical signals, physical forces, and the cytoskeleton can collaborate to generate large-scale cellular organization.