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(A) Schematic of METH CPP procedure. (B) The expression of <t>CCK1R</t> mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.
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(A) Schematic of METH CPP procedure. (B) The expression of <t>CCK1R</t> mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.
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(A) Schematic of METH CPP procedure. (B) The expression of <t>CCK1R</t> mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.
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cck antibody  (Santa Cruz Biotechnology)
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(A) Schematic of METH CPP procedure. (B) The expression of <t>CCK1R</t> mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.
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mouse polyclonal cck2r  (Santa Cruz Biotechnology)
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(A) Schematic of METH CPP procedure. (B) The expression of <t>CCK1R</t> mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.
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antibody anti cck1 receptor  (Santa Cruz Biotechnology)
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(A) Schematic of METH CPP procedure. (B) The expression of <t>CCK1R</t> mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.
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(A) Schematic of METH CPP procedure. (B) The expression of <t>CCK1R</t> mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.
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primary antibody cck1r  (Santa Cruz Biotechnology)
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(A) Schematic of METH CPP procedure. (B) The expression of <t>CCK1R</t> mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.
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(A) Schematic of METH CPP procedure. (B) The expression of CCK1R mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.

Journal: Translational Psychiatry

Article Title: CCK2R regulates METH-induced CPP acquisition within VTA-BLA-BNST circuit in male mice

doi: 10.1038/s41398-026-03982-y

Figure Lengend Snippet: (A) Schematic of METH CPP procedure. (B) The expression of CCK1R mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Membranes were then blocked for 1 h using 5% skim milk in TBS before overnight incubation at 4 °C with primary antibodies: mouse GAPDH (1:10,000 dilution, ABclonal, AC033), CCK1R (1:150, Santa Cruz, sc-514303) or CCK2R (1:150, Santa Cruz, sc-166690).

Techniques: Expressing, Western Blot, Virus, Injection, Comparison, Plasmid Preparation, In Vitro, Saline, In Vivo