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Sucrose Preference Test to Measure Anhedonic Behaviour in Mice
蔗糖偏好试验测量小鼠的快感缺乏行为   

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Neuron
Aug 2015

Abstract

The sucrose preference test (SPT) is a reward-based test, used as in indicator of anhedonia. Anhedonia, or the decreased ability to experience pleasure, represents one of the core symptoms of depression. Rodents are born with an interest in sweet foods or solutions. Reduced preference for sweet solution in SPT represents anhedonia, while this reduction can be reversed by treatment with antidepressants. SPT is carried out in the animal’s home cage. For the SPT, mice are presented with 2 dual bearing sipper tubes. One tube contains plain drinking water, and the second contains a sucrose solution. Water and sucrose solution intake is measured daily, and the positions of two bottles is switched daily to reduce any confound produced by a side bias. Sucrose preference is calculated as a percentage of the volume of sucrose intake over the total volume of fluid intake and averaged over the testing period. Here, we present our protocol that has been able to detect anhedonia in mice subjected to a chronic depression model.

Keywords: Anhedonia (快感缺失), Sucrose preference (以蔗糖偏嗜度), Chronic despair model (慢性绝望模型), Depression (抑郁)

Materials and Reagents

  1. Bearing sipper bottles
  2. Drinking bottles
  3. Laboratory-bred mice
    Note: Mice housed in groups 3-5 per cage, kept in a room with controlled temperature (~23 °C) and humidity under 12 h light/dark cycle (lights on at 7:00 AM) with ad libitum access to food and water.
  4. Sucrose (EMD Millipore, catalog number: 573113 )
  5. Plain tap water

Equipment

  1. Mouse cage without lid, which allows positioning of two bearing sipper water bottles (Figure 1)
  2. Digital balance


    Figure 1. Mouse cage with two bearing sipper bottles

Procedure

  1. Prior to beginning testing, mice are habituated to the presence of two drinking bottles for at least 3 days in their home cage.
  2. In order to detect possible side preference in the drinking behaviour, the water intake is measured daily in both bottles.
  3. After the acclimatization the mice are separated single per cage and presented to two drinking bottles: one containing 1% sucrose and the other water for 3 days in their home cage.
  4. Water and sucrose solution intake is measured daily by weighing the bottles.
  5. The positions of two bottles is switched daily to reduce any confound produced by a side bias.
  6. Sucrose preference is calculated as a percentage of the volume of sucrose intake over the total volume of fluid intake and averaged over the 3 days of testing.
    Sucrose preference = V(sucrose solution)/[V(sucrose solution)+V(water)] x 100%

Representative data



Figure 2. Sample data from sucrose preference test performed on control (Cntr), chronically despaired mice (CDM) and CDM treated for 4 weeks with classical antidepressant (CDM+AD)

Notes

  1. The SPT is an assay for anhedonia in mice subjected to a chronic depression model (Figure 2). However numerous studies show a remarkable inter-individual variability in animals’ responses to stress (Strekalova et al., 2011).
  2. Baseline sucrose preference may vary according to mouse strain and age (Pothion et al., 2004).
  3. Several factors of drinking behavior might essentially interfere with the outcome of the standard free-choice, two-bottle sucrose test paradigm:
    1. Individual mice from the same group might have a preference for drinking at one or the other corner of the cage. Housing the mice with water on both sides of the cage prior of the experiment should abolish the side preference in drinking behavior in some but not all animals. Switching the position of the bottles containing water or sucrose solution during the test will minimize this artifact (Pothion et al., 2004; Schmidt et al., 2008).
    2. The individual mice have different values of total liquid intake. The analysis of the drinking behavior should take into account the relative parameter of preference in choice paradigm, rather than of absolute intake values (Pothion et al., 2004; Strekalova and Steinbusch, 2010).
    3. The major weakness of the sucrose preference test is that mice may initially be neophobic to sucrose solution, a factor which may decrease consumption, and thus make it difficult to distinguish anhedonia from fear of an unfamiliar substance. Neophobia might be abolished by repeated exposure or habituation to sucrose solution (Strekalova and Steinbusch, 2010).
    4. Pre-exposure of mice to sucrose can increase sucrose intake and preference up to ceiling values, which decreases test’s sensitivity (Strekalova and Steinbusch, 2010). This effect can be counter balanced by application of sucrose solutions of descending concentrations (Strekalova et al., 2006).
  4. Exposing the sucrose solution to room temperature for longer than 3 days may lead to growth of bacteria and fungus, which will interfere with the final outcome of the test.
  5. Housing the mice prior to the sucrose preference test in a home cage with a water bottle placed in the middle of the cage could significantly decrease the side preference in the drinking behavior. The mice showing side preference should be excluded from the final evaluation of the results.

Acknowledgments

The protocol described here has been adapted from a previous study (Serchov et al., 2015), which succeeded in assessing anhedonia in mice subjected to model of chronic depression. This work was supported by grants from the German Research Council (DFG) (CA 115/5-4) to D.v.C. and K.B., the European Union FP7 program “MoodInflame” to D.v.C. and German Ministry for Research and Education (DMBF) grant e:bio – Modul I –ReelinSys (Project B: 031 6174A) to K.B.

References

  1. Pothion, S., Bizot, J. C., Trovero, F. and Belzung, C. (2004). Strain differences in sucrose preference and in the consequences of unpredictable chronic mild stress. Behav Brain Res 155(1): 135-146.
  2. Serchov, T., Clement, H. W., Schwarz, M. K., Iasevoli, F., Tosh, D. K., Idzko, M., Jacobson, K. A., de Bartolomeis, A., Normann, C., Biber, K. and van Calker, D. (2015). Increased signaling via adenosine A1 receptors, sleep deprivation, imipramine, and ketamine inhibit depressive-like behavior via induction of Homer1a. Neuron 87(3): 549-562.
  3. Schmidt, M. V., Sterlemann, V. and Muller, M. B. (2008). Chronic stress and individual vulnerability. Ann N Y Acad Sci 1148: 174-183.
  4. Strekalova, T., Couch, Y., Kholod, N., Boyks, M., Malin, D., Leprince, P. and Steinbusch, H. M. (2011). Update in the methodology of the chronic stress paradigm: internal control matters. Behav Brain Funct 7: 9.
  5. Strekalova, T., Gorenkova, N., Schunk, E., Dolgov, O. and Bartsch, D. (2006). Selective effects of citalopram in a mouse model of stress-induced anhedonia with a control for chronic stress. Behav Pharmacol 17(3): 271-287.
  6. Strekalova, T. and Steinbusch, H. W. (2010). Measuring behavior in mice with chronic stress depression paradigm. Prog Neuropsychopharmacol Biol Psychiatry 34(2): 348-361.

简介

蔗糖偏好测试(SPT)是基于奖励的测试,用作快感缺乏的指示器。快感缺乏或经历快乐的能力降低,代表抑郁症的核心症状之一。啮齿动物出生时对甜的食物或溶液有兴趣。 SPT中对甜味溶液的降低偏好代表了快感缺乏,而这种减少可以通过用抗抑郁药治疗逆转。 SPT在动物的家笼中进行。对于SPT,给小鼠提供2个双轴承吸管。一个管含有普通饮用水,第二个管含有蔗糖溶液。每天测量水和蔗糖溶液的摄入量,并且每天切换两个瓶的位置以减少由侧偏压产生的任何混杂。蔗糖偏好计算为蔗糖摄取量相对于流体摄入总体积的百分比,并在测试期间平均。在这里,我们提出我们的协议,已经能够检测在慢性抑郁模型小鼠中的快感。

关键字:快感缺失, 以蔗糖偏嗜度, 慢性绝望模型, 抑郁

材料和试剂

  1. 轴承吸管瓶
  2. 饮水瓶
  3. 实验室饲养的小鼠
    注意:将小鼠保持在每组3-5只,保持在具有控制温度(?23℃)和湿度的房间中,12小时光照/黑暗循环(上午7:00点亮),随意进入到食物和水。
  4. 蔗糖(EMD Millipore,目录号:573113)
  5. 纯自来水

设备

  1. 鼠笼不带盖,可以定位两个轴承吸管水瓶(图1)
  2. 数字平衡


    图1.带有两个轴承吸管瓶的鼠标笼

程序

  1. 在开始测试之前,小鼠习惯在其家笼中存在两个饮料瓶至少3天。
  2. 为了检测饮酒行为中可能的侧偏,每天在两个瓶中测量水的摄入量
  3. 在适应环境后,将小鼠每笼分离一次,并且在其笼中存在两个饮料瓶:一个含有1%蔗糖,另一个含水3天。
  4. 通过称重瓶子每天测量水和蔗糖溶液的摄入量
  5. 每天更换两个瓶子的位置以减少由侧偏压产生的任何混淆。
  6. 蔗糖偏好计算为蔗糖摄取量相对于流体摄入总体积的百分比,并且在3天的测试中平均。
    蔗糖偏好= V(蔗糖溶液)/[V(蔗糖溶液)+ V(水)]×100%

代表数据



图2.来自对照(Cntr),长期取消的小鼠(CDM)和用经典抗抑郁药(CDM + AD)治疗4周的CDM进行的蔗糖偏好测试的样品数据

笔记

  1. SPT是经历慢性抑郁模型的小鼠中的快感性测定(图2)。然而,许多研究显示动物对应激反应的显着的个体间差异(Strekalova等人,2011)。
  2. 基线蔗糖偏好可以根据小鼠品系和年龄而变化(Pothion等人,2004)。
  3. 饮酒行为的几个因素可能基本上干扰标准自由选择,两瓶蔗糖测试范例的结果:
    1. 来自同一组的个体小鼠可能喜欢在笼子的一个或另一个角上喝酒。在实验前在笼子两侧用水封住小鼠,应该在一些但不是所有动物中消除饮酒行为的偏好。在测试期间切换含有水或蔗糖溶液的瓶子的位置将最小化这种假象(Pothion等人,2004; Schmidt等人,2008)。br />
    2. 个体小鼠具有不同的总液体摄入值。饮酒行为的分析应该考虑选择范例中的偏好的相对参数,而不是绝对摄入值(Pothion等人,2004; Strekalova和Steinbusch,2010)。
    3. 蔗糖偏好试验的主要弱点是小鼠最初可能是对蔗糖溶液的疏水性,这是可能降低消耗的因素,因此使得难以区分快感缺乏与对不熟悉物质的恐惧。反复暴露或习惯于蔗糖溶液可以消除仇视恐惧症(Strekalova和Steinbusch,2010)。
    4. 小鼠预先暴露于蔗糖可以增加蔗糖摄入和偏好,直到最高值,这降低测试的敏感性(Strekalova和Steinbusch,2010)。这种效果可以通过应用递减浓度的蔗糖溶液来平衡(Strekalova等人,2006)。
  4. 将蔗糖溶液暴露于室温超过3天可能导致细菌和真菌生长,这将干扰测试的最终结果。
  5. 在家庭笼中用蔗糖偏好测试之前容纳小鼠,将水瓶放置在笼子的中间可以显着降低饮酒行为的侧偏好。显示侧偏的小鼠应该从结果的最终评价中排除。

致谢

这里描述的协议已经改编自以前的研究(Serchov等人,2015),其成功地评估在慢性抑郁模型的小鼠的快感缺乏。这项工作是由德国研究委员会(DFG)(CA 115/5-4)授予D.v.C。和K.B.,欧盟FP7程序"MoodInflame"至D.v.C.和德国研究与教育部(DMBF)给予K.B.的生物模块I-ReelinSys(项目B:031 6174A)。

参考文献

  1. Pothion,S.,Bizot,JC,Trovero,F.and Belzung,C。(2004)。  在蔗糖偏好和不可预测的慢性轻度应激的后果中的应变差异 Behav Brain Res 155(1):135-146。 />
  2. Serchov,T.,Clement,HW,Schwarz,MK,Iasevoli,F.,Tosh,DK,Idzko,M.,Jacobson,KA,de Bartolomeis,A.,Normann,C.,Biber,K.and van Calker, D.(2015)。  通过腺苷A1受体增加信号传导,睡眠剥夺,丙咪嗪和氯胺酮通过诱导Homer1a抑制抑郁样行为。 Neuron 87(3):549-562。
  3. Schmidt,MV,Sterlemann,V.and Muller,MB(2008)。  慢性压力和个人脆弱性。 Ann NY Acad Sci 1148:174-183。
  4. Strekalova,T.,Couch,Y.,Kholod,N.,Boyks,M.,Malin,D.,Leprince,P. and Steinbusch,HM(2011)。  更新慢性压力模式的方法:内部控制事务。 Behav Brain F unct 7:9.
  5. Strekalova,T.,Gorenkova,N.,Schunk,E.,Dolgov,O.和Bartsch,D.(2006)。  应用诱导的快感缺乏小鼠模型中的西酞普兰对慢性应激对照的选择性效应。 Behav Pharmacol 17 (3):271-287。
  6. Strekalova,T。和Steinbusch,HW(2010)。  测量慢性应激抑郁模式小鼠中的行为。 Prog Neuropsychopharmacol Biol Psychiatry 34(2):348-361。

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Copyright: © 2016 The Authors; exclusive licensee Bio-protocol LLC.
引用:Serchov, T., van Calker, D. and Biber, K. (2016). Sucrose Preference Test to Measure Anhedonic Behaviour in Mice. Bio-protocol 6(19): e1958. DOI: 10.21769/BioProtoc.1958.
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