Microglial Engulfment of Spines in the Ventral Zona Incerta Regulates Anxiety-Like Behaviors in a Mouse Model of Acute Pain.

Although activation of microglial cells is critical in developing brain disorders, their role in anxiety-like behaviors in pain is still vague. This study indicates that alteration of microglia's neuronal spine engulfment capacity in ventral zona incerta (ZI ) leads to significant pain and anxiety-like behaviors in mice 1-day post-injection of Complete Freud's Adjuvant (CFA1D). Performing whole-cell patch-clamp recordings in GABAergic neurons in the ZI (ZI ) in brain slices, we observed decreased activity in ZIv and reduced frequency of the miniature excitatory postsynaptic currents (mEPSCs) in ZI of CFA1D mice compared with the saline1D mice. Besides, chemogenetic activation of ZI significantly relieved pain and anxiety-like behaviors in CFA1D mice. Conversely, in naïve mice, chemogenetic inhibition of ZI induced pain and anxiety-like behaviors. Interestingly, we found changes in the density and morphology of ZI and increased microglial engulfment of spines in ZI of CFA1D mice. Furthermore, pain sensitization and anxiety-like behaviors were reversed when the ZI of CFA1D-treated mice were chemically inhibited by intra-ZI minocycline injection, accompanied by the recovery of decreased ZI excitability. Conclusively, our results provide novel insights that dysregulation of microglial engulfment capacity encodes maladaptation of ZI , thus promoting the development of anxiety-like behaviors in acute pain.