hChR2 and eNpHR3.0 viruses were used for optogenetic manipulation. hM3D(Gq) and hM4D(Gi) viruses were used for chemogenetic manipulations, Cre-dependent AAV was used to locate the PVN-projecting GABAergic neurons in BNST. (all viruses were packaged by
BrainVTA)
The viruses used in this article From BrainVTA are in the table below
|
Specific promoter |
PT-0587 rAAV-CRH-EYFP-WPRE-pA
PT-0190 rAAV-Vgat-EYFP-WPRE-pA |
|
CRE Recombinase |
PT-0136 rAAV/ retro -hSyn-Cre-WPRE-pA,
PT-0346 rAAV-Vgat-Cre-WPRE-Pa |
|
Control |
PT-0013 rAAV-Ef1α-DIO-mCherry-WPRE-pA |
|
Chemogenetics |
PT-0042 rAAV-Ef1α-DIO-hM3D(Gq)-mCherry-WPRE-pA |
|
Optogenetic |
PT-0002 rAAV-Ef1α-DIO-hChR2(H134R)-mCherry-WPRE-pA
PT-0007 rAAV-Ef1α-DIO-eNpHR3.0-mCherry-WPRE-Pa |
Pub date：2020-04-24
DOI：10.1016/j.psyneuen.2020.104690 Email：sales@brainvta.com
Yu Song, Qing-Xiang Meng, Ke Wu, Rong Hua, Zhi-Jing Song, Ying Song, Xia Qin, Jun-Li Cao, Yong-Mei Zhang
Ample evidence suggests that early life stress (ELS) is a high-risk factor for the development of visceral pain disorders, whereas the mechanism underlying neuronal circuit remains elusive. Herein, we employed neonatal colorectal distension (CRD) to induce visceral hypersensitivity in rats. A combination of electrophysiology, pharmacology, behavioral test, molecular biology, chemogenetics and optogenetics confirmed that CRD in neonatal rats could predispose the elevated firing frequency of the parvocellular corticotropin-releasing hormone (CRH) neurons in the paraventricular nucleus of hypothalamus (PVN) in adulthood, with the CRH neurons activated and the frequency of spontaneous inhibitory postsynaptic currents (sIPSC) diminished, both contributing to chronic visceral hypersensitivity. Moreover, following administration of exogenous GABA (300mM/0.5 μL) and GABA
A receptor agonist muscimol (3mM/0.5 μL) in PVN, visceral hyperalgesia was abrogated. In addition, the PVN-projecting GABAergic neurons were mainly distributed in the anterior ventral (AV) region in the bed nucleus of stria terminalis (BNST), and the excitability of these GABAergic neurons was weakened in visceral hypersensitivity. Specific depletion of the GABAergic neurons in AV region precipitated visceral hyperalgesia. Moreover, chemogenetic activation of the PVN-projecting neurons alleviated the visceral hypersensitivity. Photo activation of PVN-projecting GABAergic neurons abated the visceral hypersensitivity in neonatal-CRD rats, whereas photo inhibition evoked visceral hyperalgesia in naïve rats. Our findings demonstrated that disinhibition of the PVN-projecting GABAergic neurons in AV region contributed to the excitation of CRH neurons, thereby mediating visceral hypersensitivity. Our study might provide a novel insight into the neuronal circuits involved in the ELS-induced visceral hypersensitivity.
Fig. 1 Using virus tools to dissect PVN-projecting GABAergic neurons in AV region in BNST
To evaluate neuronal circuits involved in the ELS-induced visceral hypersensitivity, using electrophysiology, pharmacology, behavioral test, molecular biology, chemogenetics and optogenetics (From
BrainVTA) as well as pathway-specific tools，the researchers revealed a projection-specific modulation of visceral nociception and provided novel insights into the neuronal circuits and mechanisms involved in the processing of visceral pain.
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