Maternal immune activation (MIA) disrupts the central innate immune system during a critical neurodevelopmental period. Microglia are the primary innate immune cells in the brain and can mediate neurodevelopment, but the direct influence of microglia on the MIA phenotype remains largely unknown. Here, we show that MIA can lead to long-lasting effects on microglial phenotype, neuronal circuitry, and behaviors. Transcriptomic analysis revealed aberrant expression of neurogenic genes in MIA microglia. We found that microglia repopulation by colony-stimulating factor receptor 1 (CSF1R) inhibition reversed MIA-induced social deficits and corrected expression of the newly identified MIA-associated neuritogenic molecules in microglia. In vitro whole-cell patch-clamp recording and immunohistochemistry revealed that microglia repopulation restored MIA-induced changes in intrinsic excitability, dendritic spine density, and microglia-neuron interactions of layer V intrinsically bursting pyramidal neurons in the prefrontal cortex. Maternal inflammation therefore alters microglial phenotypes and changes neuronal functions by mediating microglia-neuron interactions. We found that Wingless-related MMTV integration site 5a (WNT5a) is a critical regulator of this microglia-neuron communication. Studies have shown that the neurotrophic factor WNT5a plays a critical role in neurodevelopment, and here we demonstrate that WNT5a is one of the neuritogenic genes significantly upregulated in embryonic MIA microglia. We showed using microarray analysis that the microglial secretome can promote neural stem cell differentiation through various pathways, including Wnt pathways. Live imaging of neuron-microglia co-culture demonstrated that microglia enhanced neurite development and dendritic spine density and that this was diminished by microglial Wnt5a silencing using siRNA transfection. Multi-electrode array recordings revealed that microglia co-culture increased spontaneous neuronal firing rate. Thus, microglia can secrete WNT5a and regulate dendritic spine development, maintenance, and neural circuitry. These results indicate that altered expression of microglial WNT5a due to pathogenic states such as inflammation can lead to abnormal neuronal activity. To further elucidate microglia biology, we developed an inducible immortalized murine microglial cell line using a tetracycline expression system. The addition of doxycycline can induce rapid cell proliferation for the expansion of cell colonies. Upon withdrawal of doxycycline, this monoclonal microglial cell line can differentiate and resemble in vivo microglia physiology as assessed by expression of microglial genes, innate immune response, chemotaxis, and phagocytic capabilities. This cell line becomes a convenient and useful method to study microglia in vitro. / 2024-02-03T00:00:00Z
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/43807 |
| Date | 04 February 2022 |
| Creators | Yeh, Hana |
| Contributors | Ikezu, Tsuneya |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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