Fig. 3: Nicotine-induced activation of NAc-projecting neurons triggers inhibition in the VTA. | Nature Communications

Fig. 3: Nicotine-induced activation of NAc-projecting neurons triggers inhibition in the VTA.

From: Nicotine engages a VTA-NAc feedback loop to inhibit amygdala-projecting dopamine neurons and induce anxiety-like behaviors

Fig. 3

a Retrobeads injection into the nucleus accumbens lateral shell (NAcLSh) or the basolateral amygdala (BLA). Patch-clamp recordings of DA neurons identified by tyrosine hydroxylase (TH, red), neurobiotin (NB, blue), and retrobeads (RB). Scale bar: 20 µm. b Left: Nicotine-evoked currents in NAcLSh- and BLA-projecting DA neurons by puffs at 10, 30, and 100 µM. Right: Mean current amplitudes in NAcLSh-projecting (brown: n = 8/13/11) and BLA-projecting (purple: n = 13/8/10) neurons (RM ANOVA: dose effect, F2,57 = 9.54, ***p = 0.0002; projection effect, F1,57 = 16.77, ***p = 0.0001). c Left: IPSCs and nicotinic currents before/after nicotine perfusion. Right: Mean IPSC amplitude and frequency pre- and post-nicotine in BLA-projecting (n = 14) and NAcLSh-projecting (n = 17) neurons (t-test: t30 = –2.14, *p = 0.04). d AAVretro.mCherry.Cre injection in NAc (lateral/medial shell and core) for Cre-dependent β2-nAChR re-expression in VTA. Left: TH, mCherry and GFP expression in VTA. Scale bars: 200 and 20 µm. e Firing rate responses (% change) to nicotine (30 µg/kg, black) or saline (gray) in DA neurons from GFPNAcVec (n = 19) or β2NacVec (n = 32) mice in vivo (Kolmogorov-Smirnov: β2NAcVec, D = 0.38, *p = 0.02; nicotine responses in β2NAcVec vs GFPNAcVec, D = 0.38, *p = 0.05). f Left: Mean time course of normalized firing frequency following nicotine or saline in GFPNAcVec and β2NAcVec. Right: individual firing rate variation for nicotine-activated (Nic+, n = 17, red) and inhibited (Nic-, n = 15, blue) neurons compare to baseline (paired t-test, Nic+, t16 = −4.46, ***p = 0.0004; Nic-, t14 = 3.46, **p = 0.004) and saline (paired Wilcoxon, Nic+, V = 0, ***p < 0.001; Nic-, V = 1, ***p = 0.0001). g–i Same as (d–f) with AAVretro injection into the amygdala (BLA/CeA). h GFPAmgVec (n = 10), β2AmgVec (n = 29; D = 0.45, **p = 0.005). Comparison with baseline (paired t-test, Nic+, n = 13, t12 = −6.99, ***p = 1.5e–05; Sal-, t8 = 7.52, ***p = 6.8e–05; Nic-, n = 9, t8 = 5.14, ***p = 0.0008) and saline (paired Wilcoxon, Nic+, V = 91, ***p = 0.0002; Nic-, t8 = 0.87, p = 0.4). Data are presented as mean ± SEM. All statistical tests are two-sided.

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