Disinhibition of Mesolimbic Dopamine Circuit by the Lateral Hypothalamus Regulates Pain Sensation.

Our recent study demonstrated the critical role of the mesolimbic dopamine (DA) circuit and its brain-derived neurotropic factor (BDNF) signaling in mediating neuropathic pain. The present study aims to investigate the functional role of GABAergic inputs from the lateral hypothalamus (LH) to the ventral tegmental area (VTA; LHGABA→VTA) in regulating the mesolimbic DA circuit and its BDNF signaling underlying physiological and pathologic pain. We demonstrated that optogenetic manipulation of the LHGABA→VTA projection bidirectionally regulated pain sensation in naive male mice. Optogenetic inhibition of this projection generated an analgesic effect in mice with pathologic pain induced by chronic constrictive injury (CCI) of the sciatic nerve and persistent inflammatory pain by complete Freund's adjuvant (CFA). Trans-synaptic viral tracing revealed a monosynaptic connection between LH GABAergic neurons and VTA GABAergic neurons. Functionally, in vivo calcium/neurotransmitter imaging showed an increased DA neuronal activity, decreased GABAergic neuronal activity in the VTA, and increased dopamine release in the NAc, in response to optogenetic activation of the LHGABA→VTA projection. Furthermore, repeated activation of the LHGABA→VTA projection was sufficient to increase the expression of mesolimbic BDNF protein, an effect seen in mice with neuropathic pain. Inhibition of this circuit induced a decrease in mesolimbic BDNF expression in CCI mice. Interestingly, the pain behaviors induced by activation of the LHGABA→VTA projection could be prevented by pretreatment with intra-NAc administration of ANA-12, a TrkB receptor antagonist. These results demonstrated that LHGABA→VTA projection regulated pain sensation by targeting local GABAergic interneurons to disinhibit the mesolimbic DA circuit and regulating accumbal BDNF release.SIGNIFICANCE STATEMENT The mesolimbic dopamine (DA) system and its brain-derived neurotropic factor (BDNF) signaling have been implicated in pain regulation, however, underlying mechanisms remain poorly understood. The lateral hypothalamus (LH) sends different afferent fibers into and strongly influences the function of mesolimbic DA system. Here, utilizing cell type- and projection-specific viral tracing, optogenetics, in vivo calcium and neurotransmitter imaging, our current study identified the LHGABA→VTA projection as a novel neural circuit for pain regulation, possibly by targeting the VTA GABA-ergic neurons to disinhibit mesolimbic pathway-specific DA release and BDNF signaling. This study provides a better understanding of the role of the LH and mesolimbic DA system in physiological and pathological pain.

Phenolics, Flavonoids Content and Antioxidant Activities of Tuber indicum at Different Maturity Stages.

The Chinese black truffle Tuber indicum (Ascomycota, Pezizales) is an ectomycorrhizal fungus forming hypogeous edible ascocarps. As a famous wild edible mushroom in the world, this species also attracted an increasing interest in their chemical composition and pharmacological activity. In this study, the total phenolic content, total flavonoid content and antioxidant activities of Tuber indicum collected from July to November at different maturity stages in China were analyzed. Our results showed that T. indicum collected in July (immature stage) possessed the highest amount of flavonoids (3.89 mg/g dw) and the highest ABTS.+ scavenging activity (EC50 =3.73 mg/ml). In addition, those samples collected in August (moderate mature stage) contained the highest phenolics content (4.78 mg/g dw), the highest DPPH⋅ radical scavenging activity (EC50 =3.73 mg/ml) and ferric reducing activity power (243.63 µmol FeSO4 /g). The study reveals T. indicum in the early maturity stage yield significantly higher content of phenolics and flavonoids and possessed stronger antioxidant activity than those collected in other months. This study provided important data for understanding the relationship between maturity stages and truffle formation and evaluating the quality of Chinese black truffle at different maturity.