Pu.1/Spi1 dosage controls the turnover and maintenance of microglia in zebrafish and mammals

Abstract Microglia are brain-resident macrophages playing pivotal roles in CNS development and homeostasis. Yet, the cellular and molecular basis governing microglia maintenance remain largely unknown. Here, via utilizing a visible conditional knockout allele of pu.1 gene (the master regulator for microglia/macrophage lineage development) to generate mosaic microglia populations in adult zebrafish, we show that while pu.1-deficient microglia are immediate viable, they are less competitive and chronically eliminated through Tp53-mediated cell competition. Interestingly, when conditionally inactivating Pu.1 in adult spi-b (the paralogue of zebrafish Pu.1) null mutants, microglia are rapidly depleted via apoptosis, suggesting that Pu.1 and Spi-b regulate microglia maintenance in a dosage-dependent manner. The dosage-dependent regulation of microglia maintenance by PU.1 is evolutionarily conserved in mice, as shown by conditionally inactivating single and both Pu.1 alleles in microglia respectively. Collectively, our study reveals the conserved cellular and molecular mechanisms controlling microglia turnover and maintenance in teleost and mammals.