NPY₂ Receptors Reduce Tonic Action Potential-Independent GABA_B Currents in the Basolateral Amygdala

Although NPY has potent anxiolytic actions within the BLA, selective activation of BLA NPY Y₂ receptors (Y₂Rs) acutely increases anxiety by an unknown mechanism. Using ex vivo male rat brain slice electrophysiology, we show that the selective Y₂R agonist, [ahx^5-24]NPY, reduced the frequency of GABA_A-mediated mIPSCs in BLA principal neurons (PNs). [ahx^5-24]NPY also reduced tonic activation of GABA_B receptors (GABA_BR), which increased PN excitability through inhibition of a tonic, inwardly rectifying potassium current (K ). Surprisingly, Y₂R-sensitive GABA_BR currents were action potential-independent, persisting after treatment with TTX. Additionally, the Ca²⁺-dependent, slow afterhyperpolarizing K⁺ current (I ) was enhanced in approximately half of the Y₂R-sensitive PNs, possibly from enhanced Ca²⁺ influx, permitted by reduced GABA_BR tone. In male and female mice expressing tdTomato in Y₂R-mRNA cells (tdT-Y₂R mice), immunohistochemistry revealed that BLA somatostatin interneurons express Y₂Rs, as do a significant subset of BLA PNs. In tdT-Y₂R mice, [ahx^5-24]NPY increased excitability and suppressed the K in nearly all BLA PNs independent of tdT-Y₂R fluorescence, consistent with presynaptic Y₂Rs on somatostatin interneurons mediating the above effects. However, only tdT-Y₂R-expressing PNs responded to [ahx^5-24]NPY with an enhancement of the I Ultimately, increased PN excitability via acute Y₂R activation likely correlates with enhanced BLA output, consistent with reported Y₂R-mediated anxiogenesis. Furthermore, we demonstrate the following: (1) a novel mechanism whereby activity-independent GABA release can powerfully dampen BLA neuronal excitability via postsynaptic GABA_BRs; and (2) that this tonic inhibition can be interrupted by neuromodulation, here by NPY via Y₂Rs.SIGNIFICANCE STATEMENT Within the BLA, NPY is potently anxiolytic. However, selective activation of NPY₂ receptors (Y₂Rs) increases anxiety by an unknown mechanism. We show that activation of BLA Y₂Rs decreases tonic GABA release onto BLA principal neurons, probably from Y₂R-expressing somatostatin interneurons, some of which coexpress NPY. This increases principal neuron excitability by reducing GABA_B receptor (GABA_BR)-mediated activation of G-protein-coupled, inwardly rectifying K⁺ currents. Tonic, Y₂R-sensitive GABA_BR currents unexpectedly persisted in the absence of action potential firing, revealing, to our knowledge, the first report of substantial, activity-independent GABA_BR activation. Ultimately, we provide a plausible explanation for Y₂R-mediated anxiogenesis in vivo and describe a novel and modulatable means of damping neuronal excitability.