The parabrachial nucleus (PBN) relays gustatory information as well as negative emotions. It sends dense projections to the ventral tegmental area (VTA), a heterogeneous brain region that regulates motivational behaviors. Nevertheless, the function of PBN-to-VTA connection remains elusive. To characterize the functional roles of PBN-to-VTA pathway, we first applied dual-viral retrograde tracing to specifically label VTA-projecting PBN cells. By combining c-Fos immunostainings with different behavior paradigms, we found that VTA-projecting PBN cells are significantly activated by negative affect, but not palatable feeding, thermal pain, vesicular malaise and predator threat. To further decipher the circuitry mechanisms of PBN-to-VTA input, we combined retrograde targeting approach with the Ai14 Cre reporter mouse to selectively label VTA-projecting PBN neurons and showed that PBN innervates VTA mainly through glutamatergic projection. In addition, we combined an activity-dependent targeting approach with optogenetic manipulation and found that PBN afferent activation mostly recruits VTA non-dopamine cells, which send dense projections to dorsal raphe nucleus (DR), lateral preoptic area (LPO) and peduncular part of the lateral hypothalamus (PLH), but not nucleus accumbens (NAc). Finally, to examine the behavioral functions of PBN-to-VTA input, we employed a dual-color excitatory optogenetic strategy to independently excite presynaptic input from the PBN and PBN-recruited postsynaptic VTA neurons. The results showed that both optogenetic manipulations result in aversion, dampen freely palatable feeding, and disturb instrumental food-seeking behavior in food self-administration task. Altogether, our results suggest that PBN recruits VTA non-DA cells to convey negative emotions and disengage food-seeking behavior.