Neurofibromatosis type 1-dependent alterations in mouse microglia function are not cell-intrinsic

We previously discovered a sex-by-genotype defect in microglia function using a heterozygous germline knockout mouse model of Neurofibromatosis type 1 (Nf1 ± mice), in which only microglia from male Nf1 ± mice exhibited defects in purinergic signaling. Herein, we leveraged an unbiased proteomic approach to demonstrate that male, but not female, heterozygous Nf1 ± microglia exhibit differences in protein expression, which largely reflect pathways involved in cytoskeletal organization. In keeping with these predicted defects in cytoskeletal function, only male Nf1 ± microglia had reduced process arborization and surveillance capacity. To determine whether these microglial defects were cell autonomous or reflected adaptive responses to Nf1 heterozygosity in other cells in the brain, we generated conditional microglia Nf1-mutant knockout mice by intercrossing Nf1(flox/flox) with Cx3cr1-Cre(ER) mice (Nf1(flox/wt); Cx3cr1-Cre(ER) mice, Nf1(MG) ± mice). Surprisingly, neither male nor female Nf1(MG) ± mouse microglia had impaired process arborization or surveillance capacity. In contrast, when Nf1 heterozygosity was generated in neurons, astrocytes and oligodendrocytes by intercrossing Nf1(flox/flox) with hGFAP-Cre mice (Nf1(flox/wt); hGFAP-Cre mice, Nf1(GFAP) ± mice), the microglia defects found in Nf1 ± mice were recapitulated. Collectively, these data reveal that Nf1 ± sexually dimorphic microglia abnormalities are likely not cell-intrinsic properties, but rather reflect a response to Nf1 heterozygosity in other brain cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s40478-023-01525-w.