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. 2018 Jul 9;24(1):126–144. doi: 10.1038/s41380-018-0102-9

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Effects of AdipoR1 deletion in dopamine neurons on anxiety-like behavior and VTA dopamine neuron activity. (a) Schematic diagram of generation of mice with AdipoR1 deletion in dopamine neurons. (b) Colocalization of DAT-IRES-cre (tdTomato fluorescence, red) and tyrosine hydroxylase (TH, green). (c) Real-time PCR analysis showing the Cre-mediated deletion of exon 2 of AdipoR1 in the VTA of AdipoR1^flox/flox/DAT^IREScre mice. AdipoR1^flox/flox: n = 5 mice; AdipoR1^flox/flox/DAT^IREScre: n = 4 mice. (d) Locomotor activity. AdipoR1^flox/flox: n = 14 mice; AdipoR1^flox/flox/DAT^IREScre: n = 12 mice. (e) Behavioral tests for anxiety under basal non-stress conditions. Left and middle-left, elevated plus maze test. AdipoR1^flox/flox: n = 14 mice; AdipoR1^flox/flox/DAT^IREScre: n = 11 mice. Middle-right and right, light–dark box test. AdipoR1^flox/flox: n = 14 mice; AdipoR1^flox/flox/DAT^IREScre: n = 12 mice. (f) Behavioral tests for anxiety after 30-min restraint stress. Left and middle-left, elevated plus maze test. AdipoR1^flox/flox: n = 9 mice; AdipoR1^flox/flox/DAT^IREScre: n = 7 mice. Middle-right and right, light–dark box test. AdipoR1^flox/flox: n = 8 mice; AdipoR1^flox/flox/DAT^IREScre: n = 7 mice. (g) Firing activities of VTA dopamine neurons under basal non-stress conditions. (g1) Upper, representative extracellular voltage traces from dopamine neurons in the VTA; lower, representative image demonstrating the electrode track through the VTA. (g2) Number of spontaneously active dopamine neurons per track. (g3) Left, firing rate; middle, scatter plot distribution; right, cumulative frequency distribution. (g4) Left, proportion of burst firing; middle, scatter plot distribution; right, cumulative frequency distribution. AdipoR1^flox/flox: n = 7 mice, AdipoR1^flox/flox/DAT^IREScre: n = 7 mice. (h) Firing activities of VTA dopamine neurons after 30-min restraint stress. (h1) Upper, representative extracellular voltage traces from dopamine neurons in the VTA; lower, representative image demonstrating the electrode track through the VTA. (h2) Number of spontaneously active dopamine neurons per track. (h3) Left, firing rate; middle, scatter plot distribution; right, cumulative frequency distribution. (h4) Left, proportion of burst firing; middle, scatter plot distribution; right, cumulative frequency distribution. AdipoR1^flox/flox: n = 7 mice, AdipoR1^flox/flox/DAT^IREScre: n = 7 mice. *P < 0.05, **P < 0.01 compared with AdipoR1^flox/flox littermate controls

Fig. 4 — Effects of AdipoR1 deletion in dopamine neurons on anxiety-like behavior and VTA dopamine neuron activity. (a) Schematic diagram of generation of mice with AdipoR1 deletion in dopamine neurons. (b) Colocalization of DAT-IRES-cre (tdTomato fluorescence, red) and tyrosine hydroxylase (TH, green). (c) Real-time PCR analysis showing the Cre-mediated deletion of exon 2 of AdipoR1 in the VTA of AdipoR1^flox/flox/DAT^IREScre mice. AdipoR1^flox/flox: n = 5 mice; AdipoR1^flox/flox/DAT^IREScre: n = 4 mice. (d) Locomotor activity. AdipoR1^flox/flox: n = 14 mice; AdipoR1^flox/flox/DAT^IREScre: n = 12 mice. (e) Behavioral tests for anxiety under basal non-stress conditions. Left and middle-left, elevated plus maze test. AdipoR1^flox/flox: n = 14 mice; AdipoR1^flox/flox/DAT^IREScre: n = 11 mice. Middle-right and right, light–dark box test. AdipoR1^flox/flox: n = 14 mice; AdipoR1^flox/flox/DAT^IREScre: n = 12 mice. (f) Behavioral tests for anxiety after 30-min restraint stress. Left and middle-left, elevated plus maze test. AdipoR1^flox/flox: n = 9 mice; AdipoR1^flox/flox/DAT^IREScre: n = 7 mice. Middle-right and right, light–dark box test. AdipoR1^flox/flox: n = 8 mice; AdipoR1^flox/flox/DAT^IREScre: n = 7 mice. (g) Firing activities of VTA dopamine neurons under basal non-stress conditions. (g1) Upper, representative extracellular voltage traces from dopamine neurons in the VTA; lower, representative image demonstrating the electrode track through the VTA. (g2) Number of spontaneously active dopamine neurons per track. (g3) Left, firing rate; middle, scatter plot distribution; right, cumulative frequency distribution. (g4) Left, proportion of burst firing; middle, scatter plot distribution; right, cumulative frequency distribution. AdipoR1^flox/flox: n = 7 mice, AdipoR1^flox/flox/DAT^IREScre: n = 7 mice. (h) Firing activities of VTA dopamine neurons after 30-min restraint stress. (h1) Upper, representative extracellular voltage traces from dopamine neurons in the VTA; lower, representative image demonstrating the electrode track through the VTA. (h2) Number of spontaneously active dopamine neurons per track. (h3) Left, firing rate; middle, scatter plot distribution; right, cumulative frequency distribution. (h4) Left, proportion of burst firing; middle, scatter plot distribution; right, cumulative frequency distribution. AdipoR1^flox/flox: n = 7 mice, AdipoR1^flox/flox/DAT^IREScre: n = 7 mice. *P < 0.05, **P < 0.01 compared with AdipoR1^flox/flox littermate controls