Microglial MyD88-dependent pathways are regulated in a sex specific manner in the context of HMGB1-induced anxiety

Chronic stress is a major risk factor for development and recurrence of anxiety disorders. Chronic stress has been shown to impact the immune system, causing microglial activation in the medial prefrontal cortex (mPFC), a brain region involved in the pathogenesis of anxiety. HMGB1 is both an established modulator of neuronal firing and a potent pro-inflammatory stimulus that is released from neuronal and non-neuronal cells following stress. HMGB1 in the context of stress acts as a danger associated molecular pattern (DAMP) instigating robust proinflammatory responses throughout the brain; so much so, that localized drug delivery of HMGB1 alters behavior in the absence of any other forms of stress i.e. social isolation, or behavioral stress models. Few studies have investigated the molecular mechanisms which underlie HMGB1 associated behavioral effects in a cell-specific manner. The aim of this study is to investigate cellular and molecular mechanisms underlying HMGB1 induced behavioral dysfunction with regards to cell-type specificity and potential sex differences. Here, we report that both male and female mice exhibited anxiety-like behavior following increased HMGB1 in the mPFC as well as changes in microglial morphology. However, only female mice showed microglial activation characterized by increased phagocytic capacity and decreased Tmem119 expression. Moreover, when measuring RNA from isolated microglia and non-microglial cells from the frontal cortex we found that female HMGB1 treated mice displayed a robust increase of RAGE and MyD88. Given these findings, to further investigate the underlying molecular mechanisms associated with HMGB1 induced anxiety-like behavior and microglia activation in mice, we used cKO (conditional knockout) mice with conditional deletion of MyD88 in microglia and repeated the paradigm. For males, we saw that the genetic manipulation did not prevent behavioral deficits in response to HMGB1 treatment. However, female cKO mice were protected from HMGB1-induced behavioral deficits. This study supports the hypothesis that the MyD88 signaling in microglia may be a crucial mediator of stress response in adult female mice.