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Sucrose preference test for measurement of stress-induced anhedonia in mice

Nature Protocols volume 13, pages 1686–1698 (2018)Cite this article

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Abstract

Anhedonia is the inability to experience pleasure from rewarding or enjoyable activities and is a core symptom of depression in humans. Here, we describe a protocol for the measurement of anhedonia in mice, in which anhedonia is measured by a sucrose preference test (SPT) based on a two-bottle choice paradigm. A reduction in the sucrose preference ratio in experimental relative to control mice is indicative of anhedonia. To date, inconsistent and variable results have been reported following the use of the SPT by different groups, probably due to the use of different protocols and equipment. In this protocol, we describe how to set up a clearly defined apparatus for SPT and provide a detailed protocol to ensure greater consistency when carrying out SPT. This optimized protocol is highly sensitive, reliable, and adaptable for evaluation of chronic stress–related anhedonia, as well as morphine-induced dependence. The whole SPT, including adaptation, baseline measurement, and testing, takes 8 d.

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Fig. 1: Design of sucrose preference test apparatus.
Fig. 2: Example of the use of SPT to measure stress-induced anhedonia.
Fig. 3: Example of SPT to measure the effects of an addictive drug.

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Acknowledgements

This research was supported by grants from the National Key Research and Development Program of China (2016YFC1306703 to D.-Y.Z.), the National Natural Science Foundation of China (81571269 to Q.-G.Z., 31530091 to D.-Y.Z., 31771334 to H.C., and 31671107 to L.-J.Z.), the Major Research Plan of the National Natural Science Foundation of China (91649125 to H.C.), and the Natural Science Foundation of Jiangsu Province (BK20170021 to L.-J.Z.). This behavioral procedure was adapted from previously published studies7 and was improved and performed by our group as described12,22,34. This study was also supported by the Key Lab of Cardiovascular and Cerebrovascular Drugs of Jiangsu Province and by the Collaborative Innovation Center For Cardiovascular Disease Translational Medicine for data collection, analysis, and interpretation.

Author information

Author notes

  1. These authors contributed equally: Meng-Ying Liu, Chun-Yu Yin, and Li-Juan Zhu.

Authors and Affiliations

  1. Department of Clinical Pharmacology, Pharmacy College, Nanjing Medical University, Nanjing, China

    Meng-Ying Liu, Chun-Yu Yin, Li-Juan Zhu, Xian-Hui Zhu, Chu Xu, Chun-Xia Luo, Dong-Ya Zhu & Qi-Gang Zhou

  2. Department of Pharmacy, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, China

    Meng-Ying Liu

  3. Key Laboratory of Developmental Genes and Human Diseases, MOE, Department of Histology and Embryology, School of Medicine, Southeast University, Nanjing, China

    Li-Juan Zhu

  4. Key Laboratory of Cardiovascular & Cerebrovascular Medicine, School of Pharmacy, Nanjing Medical University, Nanjing, China

    Hongshan Chen

  5. Department of Cardiothoracic Surgery, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, China

    Hongshan Chen

  6. Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing, China

    Hongshan Chen

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Contributions

M.-Y.L., C.-Y.Y., L.-J.Z., C.X. and X.-H.Z. acquired the data. C.-X.L. gave advice for revision of the article. D.-Y.Z. and H.C. provided materials for the experiments. Q.-G.Z., D.-Y.Z., and H.C. prepared and wrote the article.

Corresponding authors

Correspondence to Hongshan Chen, Dong-Ya Zhu or Qi-Gang Zhou.

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Related links

Key references using this protocol:

Zhou, Q. G. et al. J. Neurosci. 31, 7579–7590 (2011): https://doi.org/10.1523/JNEUROSCI.0004-11.2011

Zhou, Q. G. et al. J. Neurosci. 31, 12258–12269 (2011): https://doi.org/10.1523/JNEUROSCI.0805-11.2011

Zhou, Q. G. et al. Transl. Psychiatry. 6, e836 (2016): https://doi.org/10.1038/tp.2016.106

Integrated supplementary information

Supplementary Figure 1 The weight of mice is altered by CMS and fluoxetine.

(a) The weight of mice exposed to 28-day CMS is lower than those of control mice. n = 15. (b) Exposure to 56-day CMS led to decreased weight of mice, reversed by fluoxetine treatment. n = 15. *P < 0.01, **P < 0.01, ***P < 0.001. Student’s t test for a. One-way ANOVA for b. Error bars, s.e.m. All the experiments were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University

Supplementary Figure 2 Associated intakes of sucrose and regular water solutions altered by CMS and fluoxetine.

(a) The intakes of sucrose and regular water solutions in SPT before CMS. n = 11 – 12 for day 1, n = 12 – 14 for day 2, and n = 14 – 15 in each group for test. (b) The intakes of sucrose and regular water solutions in SPT after 28 days CMS exposure. n = 12-14 for day 1, n = 14 for day 2, and n = 15 in each group for test. (c) The intakes of sucrose and regular water solutions in SPT after 56 days CMS exposure with or without fluoxetine treatment during day 22 – 56. n = 12-13 for day 1, n = 13 – 14 for day 2, and n = 13 – 15 in each group for test. Invalid data caused by several conditions including no drinking, over drinking, and no preference was excluded in according to the criteria described in the protocol. Error bars, s.e.m. All the experiments were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University

Supplementary Figure 3 Associated intakes of sucrose and regular water solutions altered by CSDS.

(a) The intakes of sucrose and regular water solutions in SPT after CSDS. n = 12 – 13 for day 1, n = 13 – 14 for day 2, n = 14 for test. Invalid data caused by several conditions including no drinking, over drinking, and no preference was excluded in according to the criteria described in the protocol. Error bars, s.e.m. All the experiments were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University

Supplementary Figure 4 Associated intakes of sucrose and regular water solutions altered by morphine.

(a) The intakes of sucrose and regular water solutions in SPT after morphine dependence. n = 11 – 12 for day 1, n = 12 – 14 for day 2, n = 15 for test. (b) The intakes of sucrose and regular water solutions in SPT after morphine withdrawal. n = 12 – 14 for day 1, n = 15 for day 2, n = 15 for test. Error bars, s.e.m. Invalid data caused by several conditions including no drinking, over drinking, and no preference was excluded in according to the criteria described in the protocol. All the experiments were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University

Supplementary Figure 5 Global withdrawal score after morphine withdrawal.

(a) The global withdrawal score of mice treated with saline or morphine. Withdrawal was precipitated in both groups by the administration of the opioid receptor antagonist, naloxone (1 mg/kg, s.c.). Somatic signs of withdrawal, including jumping, wet dog shakes, paw tremor, teeth chattering, body tremor, ptosis, piloerection, and sniffing, were recorded for 30 minutes immediately after administration. The global withdrawal score was calculated for each animal by giving a relative weight to each withdrawal sign. n = 15 in each group. ***P < 0.001. Student’s t test. Error bars, s.e.m. All the experiments were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University

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Liu, MY., Yin, CY., Zhu, LJ. et al. Sucrose preference test for measurement of stress-induced anhedonia in mice. Nat Protoc 13, 1686–1698 (2018). https://doi.org/10.1038/s41596-018-0011-z

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