Glutamatergic Projection from the Ventral Tegmental Area to the Zona Incerta Regulates Fear Response

Innate defensive behavior is important for animal survival. The Vglut²⁺ neurons in the ventral tegmental area (VTA) have been demonstrated to play important roles in innate defensive behaviors, but the neural circuit mechanism is still unclear. Here, we find that VTA - zona incerta (ZI) glutamatergic projection is involved in regulating innate fear responses. Combining calcium signal recording and chemogentics, we find that VTA-Vglut²⁺ neurons respond to foot shock stimulus. Inhibition of VTA-Vglut²⁺ neurons reduces foot shock-evoked freezing, while chemogentic activation of these neurons results in an enhanced fear response. Using viral tracing and immunofluorescence, we show that VTA - Vglut²⁺ neurons send direct excitatory outputs to the ZI. Moreover, we find that the activity of VTA^Vglut2 - ZI projection is pivotal in modulating fear response. Together, our study reveals a new VTA - ZI glutamatergic circuit in mediating innate fear response and provides a potential target for treating post-traumatic stress disorder.