A unique population of ventral tegmental area (VTA) neurons co-transmits glutamate and GABA. However, the circuit inputs to VTA VGluT2+VGaT+ neurons are unknown, limiting our understanding of their functional capabilities. By coupling monosynaptic rabies tracing with intersectional genetic targeting in male and female mice, we found that VTA VGluT2+VGaT+ neurons received diverse brainwide inputs. The largest numbers of monosynaptic inputs to VTA VGluT2+VGaT+ neurons were from superior colliculus (SC), lateral hypothalamus (LH), midbrain reticular nucleus, and periaqueductal gray, whereas the densest inputs relative to brain region volume were from the dorsal raphe nucleus, lateral habenula, and VTA. Based on these and prior data, we hypothesized that LH and SC inputs were from glutamatergic neurons. Optical activation of glutamatergic LH neurons activated VTA VGluT2+VGaT+ neurons regardless of stimulation frequency and resulted in flee-like ambulatory behavior. In contrast, optical activation of glutamatergic SC neurons activated VTA VGluT2+VGaT+ neurons for a brief period of time at high frequency and resulted in head rotation and arrested ambulatory behavior (freezing). Stimulation of glutamatergic LH neurons, but not glutamatergic SC neurons, was associated with VTA VGluT2+VGaT+ footshock-induced activity and inhibition of LH glutamatergic neurons disrupted VTA VGluT2+VGaT+ tailshock-induced activity. We interpret these results such that inputs to VTA VGluT2+VGaT+ neurons may integrate diverse signals related to the detection and processing of motivationally salient outcomes.

一个独特的腹侧被盖区 （VTA） 神经元群共同传递谷氨酸和 GABA。然而，VTA VGluT2+VGaT+ 神经元的电路输入是未知的，这限制了我们对它们功能的理解。通过将单突触狂犬病追踪与雄性和雌性小鼠的交叉遗传靶向相结合，我们发现 VTA VGluT2+VGaT+ 神经元接受了不同的全脑输入。VTA VGluT2+VGaT+ 神经元的最大单突触输入来自上丘 （SC）、下丘脑外侧 （LH）、中脑网状核和导水管周围灰质，而相对于脑区体积最密集的输入来自中缝背核、外侧缰绳和 VTA。根据这些和先前的数据，我们假设 LH 和 SC 输入来自谷氨酸能神经元。谷氨酸能 LH 神经元的光学激活了 VTA VGluT2+VGaT+ 神经元，无论刺激频率如何，都导致了逃跑样的走动行为。相比之下，谷氨酸能 SC 神经元的光学激活在短时间内高频激活了 VTA VGluT2+VGaT+ 神经元，并导致头部旋转和行走行为停止（冻结）。刺激谷氨酸能 LH 神经元，而不是谷氨酸能 SC 神经元，与 VTA VGluT2+VGaT+ 足震诱导的活动相关，而抑制 LH 谷氨酸能神经元破坏了 VTA VGluT2+VGaT+ 尾震诱导的活动。我们解释这些结果，以便 VTA VGluT2+VGaT+ 神经元的输入可以整合与动机显着结果的检测和处理相关的不同信号。