一组老鼠表现出强迫性吸食可卡因的行为。(A)鉴定强迫性可卡因使用大鼠的实验时间表。(B)基于惩罚阶段2和3 (P2和P3)参数的t分布随机邻居嵌入(t-SNE)投影和包括抗性(集群1)和敏感(集群2)大鼠的集群t-SNE三维表示的分层聚类。行为表现包括主动刺鼻(A)，不主动刺鼻(IA)，无效刺鼻和可卡因注射。(C)敏感大鼠和抵抗大鼠的比例(左)，以及基线和第三次惩罚阶段的可卡因注射(右)。与耐药大鼠相比，敏感大鼠输注较少。双向方差分析(ANOVA)显示交互作用有统计学意义，F1,89 = 80.76, P < 0.001，事后分析，P3敏感组与耐药组***P < 0.001, P3敏感组与耐药组与基线组###P < 0.001;N = 60和31。(D和E)从获取过程中获得的积极的、不活跃的和可卡因注射的鼻塞数量。敏感鼠和抵抗鼠之间没有差异; two-way ANOVA showed nonsignificant main effects on groups, F1,89 = 0.53, P = 0.4671 for active nosepokes, F1,89 = 0.03, P = 0.8639 for inactive nosepokes, and F1,89 = 0.14, P = 0.7086 for infusions; n = 60 and 31, respectively. NS, not significant. Data are presented as mean values ± SEMs. Credit:科学的进步(2022)。DOI: 10.1126 / sciadv.abq5745
抗休克大鼠aIC神经元过度活跃。(A)第三次惩罚测试后90分钟大脑区域c-Fos的映射。OFC的未配对t检验，t11 = 2.24， *P = 0.046;t10 = 4.21， **P = 0.005;NAc岩心t12 = 2.50， *P = 0.039;NAc壳t12 = 2.53， *P = 0.027;t12 = 4.13， **P = 0.004为DLS;t12 = 2.71， *P = 0.03;N = 5 ~ 6, 7 ~ 8。(B) aIC c-Fos免疫荧光染色的代表性图像。 Scale bars, 500 μm (left) and 50 μm (right). (C) Schematic of calcium photometry. Scale bar, 1 mm. (D and E) Heatmap and plot of z-scored aIC Ca2+ signals in no punish, sensitive, and resistant group; n = 5 for each group. (F) AUC of z-scored aIC Ca2+ signals in 0 to 2 s. One-way ANOVA, F2,12 = 17.58, P < 0.001, post hoc analysis, *P = 0.028 for sensitive versus resistant, ***P < 0.001 for no punish versus resistant. Number of events per minute of aIC neurons. One-way ANOVA, F2,12 = 36.00, P < 0.001, post hoc analysis, ***P < 0.001 for sensitive versus resistant and no punish versus resistant; n = 5 for each group. (G) Amplitude and frequency of sEPSCs of aIC neurons. Unpaired t test, t32 = 2.12, *P = 0.042 and t32 = 3.60, **P = 0.001, respectively; 13 neurons from 3 sensitive rats and 21 neurons from 5 resistant rats. Cg1, cingulate cortex area 1; Cg2, cingulate cortex area 2; PrL, prelimbic cortex; IL, infralimbic cortex; NAc core/shell, nucleus accumbens core/shell; MS, medial septum; DLS, dorsolateral striatum; BLA, basolateral amygdala; CeA, central amygdala; PVT, paraventricular thalamus; LHb, lateral habenula. Data are presented as mean values ± SEMs. Credit:科学的进步， DOI: 10.1126/sciadv.abq5745
aIC神经元的激活对于强迫性可卡因使用是必要的，也是足够的。(A)病毒输注的实验时间表和示意图，以分析抑制aIC后耐药大鼠的强迫性可卡因使用行为。(B) AAV-mCherry组和AAV-hM4D组的可卡因注射液。双因素方差分析，F1,18 = 22.51, P < 0.001，事后分析，P4, P5, P6 ***P < 0.001;每组N = 10。(C) aIC抑制前后AAV-mCherry组和AAV-hM4D组大鼠敏感和耐药比例;Fisher精确检验，***P < 0.001。(D)病毒输注的实验时间表和示意图，分析aIC激活后的强迫性可卡因使用行为。鳞片条，1mm。(E)可卡因输注从AAV-mCherry和AAV-hM3D组获得。双向方差分析，F1,25 = 15.22, P < 0.001，事后分析，P2和P3的***P < 0.001; n = 14 and 13, respectively. (F) Ratio of sensitive and resistant rats in AAV-mCherry and AAV-hM3D group after aIC activation; Fisher's exact test, **P = 0.004. Data are presented as mean values ± SEMs. Credit:科学的进步， DOI: 10.1126/sciadv.abq5745
aIC谷氨酸神经元双向调节强迫性可卡因使用。(A)病毒输注的实验时间表和示意图，以分析耐药大鼠aIC谷氨酸神经元抑制后的强迫性可卡因使用行为。(B) AAV-mCherry组和AAV-hM4D组的可卡因注射液。双因素方差分析，F1,16 = 9.89, P = 0.006，事后分析，P4 **P = 0.001, P5、P6 ***P < 0.001;N = 10和8。(C) AAV-mCherry组和AAV-hM4D组大鼠抑制aIC谷氨酸能神经元前后敏感和抵抗的比例;Fisher精确检验，***P < 0.001。(D)病毒输注的实验时间线和示意图，以分析aIC谷氨酸神经元激活后的强迫性可卡因使用行为。鳞片条，1mm。(E) AAV-mCherry组和AAV-hM3D组的可卡因注射液。双因素方差分析，F1,24 = 6.42, P = 0.018，事后分析，P2 **P = 0.004, P3 ***P < 0.001; n = 12 and 14, respectively. (F) Ratio of sensitive and resistant rats in AAV-mCherry and AAV-hM3D group after activation of aIC glutamatergic neurons; Fisher's exact test, *P = 0.014. Data are presented as mean values ± SEMs. Credit:科学的进步， DOI: 10.1126/sciadv.abq5745
在耐药大鼠中，从OFC到aIC的谷氨酸投射增强。(A)接受OFC输入的aIC神经元的全细胞膜片钳记录的实验时间轴和示意图。比例尺，左为1mm，右为100 μm。(B) OFC-aIC神经元记录的sEPSCs振幅和频率。无配对t检验，振幅t35 = 2.42， *P = 0.021;t35 = 10.71，频率***P < 0.001, 21个神经元来自4只敏感大鼠，16个神经元来自4只抗性大鼠。(C)从接受OFC输入的aIC神经元获得的响应增加阶跃电流的ap数量。双因素方差分析，f1187 = 297.4, P < 0.001，事后分析，200 ~ 600 pA *P < 0.05, 4只敏感大鼠18个神经元，3只抗性大鼠19个神经元。(D)全细胞膜片钳记录aIC神经元对470纳米激光反应的实验时间线和示意图(2ms，蓝色条)。(E和F)与不惩罚和敏感大鼠相比，抗性大鼠aIC神经元PPR显著降低。 One-way ANOVA, F2,45 = 4.66, P = 0.0145, post hoc analysis, *P = 0.0375 for sensitive versus resistant, and *P = 0.0199 for no punish versus resistant. (G and H) The A/N ratio of aIC neurons was significantly increased in resistant rats. One-way ANOVA, F2,45 = 54.36, P < 0.001, post hoc analysis, ***P < 0.001 for sensitive versus resistant and no punish versus resistant; 16 neurons from three no punish rats, 18 neurons from three sensitive rats, and 14 neurons from three resistant rats. Data are presented as mean values ± SEMs. Credit:科学的进步， DOI: 10.1126/sciadv.abq5745
