Training session, mice had been run each day at each photostimulation frequency (1, 5, 10, 20, 40 Hz) within a counter-balanced design. Optical self-inhibition of VgatVTA::NpHR neurons VgatVTA::NpHR and VgatVTA::Handle mice with optical fibers implanted above the VTA had been trained in 1 30 min session to nose poke on a fixed ratio (FR-1) schedule for photoinhibition of VTA GABAergic cell bodies in typical mouse operant chambers as described above (Med Associates). Photostimulation of VgatBNSTvVTA::ChR2 projections and photoinhibition of VgatVTA::NpHR neurons throughout the elevated plus maze VgatBNSTvVTA::ChR2, VgatVTA::NpHR, VgatVTA::Manage, and VgatBNSTvVTA::Manage mice have been run in the elevated plus maze (EPM) to assay anxiety-like behavior. Activity and location was recorded for 5 min (baseline).1206981-68-1 site Following this five min baseline period, VgatBNSTvVTA::ChR2 and VgatBNSTvVTA::Manage mice received continuous 20 Hz photostimulation for five min, when VgatVTA::NpHR and VgatVTA::Manage mice received continual inhibition for five min. Quickly following the five min photostimulation or photoinhibition epoch, all mice had a 5 min period in which they received no light delivery. Photostimulation of VgatBNSTvVTA::ChR2 projections for the duration of foot-shock followed by freezing and anxiety-like behavior measurements VgatBNSTvVTA::ChR2 and VgatBNSTvVTA::Control mice with optical fibers implanted above the VTA were run within a modified foot-shock paradigm as described above. Briefly, mice have been placed into sound attenuated mouse chambers (Med Associates) to get a 5 min baseline period. Following the 5 min baseline period, a home light and white noise were activated and mice received the same foot shock protocol as described above. Additionally, throughout the 20 min shock session, all mice received constant 20 Hz photostimulation. A separate cohort of mice (VgatBNSTvVTA::ChR2 and VgatBNSTvVTA::Manage) received constant 20 Hz photostimulation of this pathway within the absence of foot shock. Promptly following the 20 min foot shock and photostimulation epoch, all mice had a five min period in which they received no foot shock or photostimulation while nevertheless exposed to contextual cues, to assay freezing behavior. Freezing was defined because the total lack of any movement, except respiration for a period of two s. The 30 min test session was recorded using a CCD camera that was interfaced with Ethovision application (Noldus Data Technologies).Price of 4-Formyl-3-hydroxybenzoic acid Time frozen through the 5 min period straight away following the foot shock and photostimulation session was recorded.PMID:24883330 Roughly three hr following the foot shock and photostimulation session or simply the photostimulation session inside the absence of foot shock, mice have been run on the elevated-plus maze to assay anxiety-like behavior for five min.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptSupplementary MaterialRefer to Net version on PubMed Central for supplementary material.Nature. Author manuscript; accessible in PMC 2013 October 11.Jennings et al.PageAcknowledgmentsWe thank Malhar Patel, Jana Phillips, Scot Maciver, for help, Dr. Vladimir Gukassyan as well as the UNC Neuroscience Center Microscopy Core (P30 NS045892), and members from the Stuber lab for discussion. We thank Dr. Karl Deisseroth for viral constructs and the UNC vector core facility for viral packaging. We thank Drs. Bradford Lowell and Linh Vong for supplying the Vgat-ires-cre and Vglut2-ires-cre mice. This study was supported by The Whitehall Foundation, The Foundation of Hope, and.