title: Silencing the NR2B gene in rat ACC neurons by lentivirus-delivered shRNA alleviates pain-related aversion.
abstract: The N-methyl-D-aspartate (NMDA) receptor NR2B subunit on neurons in the anterior cingulate cortex (ACC) is implicated in the affective response to noxious stimuli. Selectively silencing this NR2B subunit in ACC neurons could therefore alleviate pain-related aversion. However, to date, there is no optimal approach to selectively silence the NR2B gene in ACC neurons. In the present study, we constructed lentiviral vectors and delivered shRNA (NR2B-RNAi-LV) to effectively silence the NR2B gene in ACC neurons. The use of lentivirus resulted in 95% transfection efficiency and 83% silencing of the NR2B gene in ACC neurons. Electrophysiological experiments showed that the total INMDA was similarly reduced by 48% in lentivirus-transfected ACC neurons. The biochemical and functional data demonstrated that lentiviral shRNA delivery produced a high transfection and silencing efficiency in the ACC neurons. SNI rats weighting 220-250 g were randomly divided into three groups: normal saline group (NS), lenti-siRNA/NC (LV-NC) group, and lenti-siRNA/NR2B (LV-NR2B) group, and conditioned place avoidance was conducted. The results indicated that NR2B-RNAi-LV decreased greatly the conditioning scores of F-CPA while NC-GFP-LV has no effects. NR2B mRNA expression in the NR2B-RNAi-LV group was significantly lower than that in the control group and NC-GFP-LV group. This novel approach of silencing the NR2B gene in ACC neuron could potentially be used to alleviate pain-related aversion.
authors.affiliation: Department of Neurology, Shandong Provincal Hospital Affiliated to Shandong University Jinan 250021, China.
authors.forename: Shou-Gang
authors.initials: SG
authors.lastname: Guo
authors.affiliation: Department of Nephrology, Taian Central Hospital Taian 271000, China.
authors.forename: Xiu-Hua
authors.initials: XH
authors.lastname: Lv
authors.affiliation: Department of Nephrology, Taian Central Hospital Taian 271000, China.
authors.forename: Shan-Hui
authors.initials: SH
authors.lastname: Guan
authors.affiliation: Department of Anesthesiology, Shandong Provincal Hospital Affiliated to Shandong University Jinan 250021, China.
authors.forename: Hui-Lu
authors.initials: HL
authors.lastname: Li
authors.affiliation: Department of Anesthesiology, The Second Hospital of Shandong University Jinan 250021, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Qiao
authors.affiliation: Department of Anesthesiology, The Second Hospital of Shandong University Jinan 250021, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Feng
authors.affiliation: Department of Neurology, Shandong Provincal Hospital Affiliated to Shandong University Jinan 250021, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Cong
authors.affiliation: Department of Anesthesiology, Shandong Provincal Hospital Affiliated to Shandong University Jinan 250021, China.
authors.forename: Gong-Ming
authors.initials: GM
authors.lastname: Wang
keywords: ACC; NR2B; RNA interference; gene silencing