Tion connected to aversive stimuli and their connected contextual cues. Electrical stimulation on the BNST produces each excitatory and inhibitory responses in VTA neurons in vivo6 implying that distinct subcircuits may perhaps exist. In mice injected with AAV5-CaMKIIa-ChR2-eYFP to nonspecifically target BNSTvVTA projection neurons (CaMKIIaBNSTvVTA::ChR2), whole-cell recordings in brain slices revealed that photostimulation on the CaMKIIaBNSTvVTA pathway developed each glutamatergic and aminobutyric acid (GABAergic) currents in VTA neurons (Supplementary Fig. 4), demonstrating that neurochemically distinct BNSTv neurons project towards the VTA. We next dissected the functional connectivity between distinct glutamatergic and GABAergic BNSTvVTA neurons and their genetically defined postsynaptic targets within the VTA. Injection of a cre-inducible viral construct coding for ChR2-eYFP into the BNSTv in Vglut2-ires-cre or Vgat-ires-cre mouse lines26 resulted in robust expression in the BNSTv at the same time as in fibers originating from these neurons that innervated the VTA (Fig. 2a,b). Whole-cell recordings from VTA neurons revealed that photostimulation of ChR2containing fibers originating from Vglut2 (Vglut2BNSTvVTA::ChR2) or Vgat (VgatBNSTvVTA::ChR2) expressing BNSTv neurons made excitatory or inhibitory postsynaptic currents respectively (Fig. 2c,d; Supplementary Fig. five). Vglut2BNSTvVTA and VgatBNSTvVTA terminals formed functional synapses mostly onto non-dopaminergic and medially located dopaminergic neurons, which have already been implicated in responding to aversive stimuli7,9,11,13 (Fig.1083246-26-7 Chemical name 2e,f; Supplementary Fig 6?; Supplementary Strategies). These information deliver a circuit blueprint by which BNSTv subcircuits interface with VTA-reward circuitry. We subsequent explored irrespective of whether glutamatergic or GABAergic subpopulations of BNSTvVTA neurons differentially respond to foot-shock sessions and linked contextual cues. Employing optical antidromic activation in vivo, we identified 34/145 Vglut2BNSTvVTA::ChR2 expressing neurons (Supplementary Fig. 1; Supplementary Fig. 8; Supplementary Table 1). Though all projection neurons displayed heterogeneous firing patterns (Supplementary Fig. three), identified Vglut2BNSTvVTA projection neurons exhibited a net enhancement of firing throughout the aversive occasion (Fig. 3a,b). In contrast, 33/77 identified VgatBNSTvVTA::ChR2 expressing neurons principally exhibited reduced firing in the course of the aversive event (Fig.3-Butynoic acid Chemscene 3c,d; Supplementary Fig.PMID:35850484 1; Supplementary Fig. 8; Supplementary Table 1). Furthermore, 1 week immediately after five consecutive everyday foot-shock sessions, re-exposure to shock-associated contextual cues alone resulted within a net enhancement of Vglut2BNSTvVTA::ChR2 neuronal activity (Fig. 3e,f; Supplementary Fig. 9) while the activity of VgatBNSTvVTA::ChR2 neurons had been largely suppressed (Fig. 3g,h; Supplementary Fig. 9). Collectively, exposure for the aversive event or associated cues alone enhanced the firing of Vglut2BNSTvVTA neurons, though simultaneously suppressing the activity of VgatBNSTvVTA neurons.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptNature. Author manuscript; readily available in PMC 2013 October 11.Jennings et al.PageSince aversive stimuli enhanced the activity of Vglut2BNSTvVTA neurons (Fig. 3a,b,e,f), which can excite non-dopaminergic VTA neurons (Fig. 2e), we subsequent explored the behavioral consequences of selectively activating this projection in behaving mice. We tested mice inside a real-time location preference paradigm to as.