Martine Hascoët

The Mouse Light–Dark Box Test

The light/dark test is based on the innate aversion of rodents to brightly illuminated areas and on the spontaneous exploratory behaviour of rodents in response to mild stressors, that is, novel environment and light. The test apparatus consists of a small dark safe compartment (one third) and a large illuminated aversive compartment (two thirds). The test was developed with male mice. The strain, weight and age may be crucial factors. The extent to which an anxiolytic compound can facilitate exploratory activity depends on the baseline level in the control group. Differences between the type and severity of external stressors might account for the variable results reported by different laboratories. The light/dark test may be useful to predict anxiolytic-like or anxiogenic-like activity in mice. Transitions have been reported to be an index of activity-exploration because of habituation over time, and the time spent in each compartment to be a reflection of aversion. Classic anxiolytics (benzodiazepines) as well as the newer anxiolytic-like compounds (e.g. serotonergic drugs or drugs acting on neuropeptide receptors) can be detected using this paradigm. It has the advantages of being quick and easy to use, without requiring the prior training of animals.

The mouse ligth-dark paradigm: A review

1. The light/dark paradigm is based on the innate aversion of rodents to brightly illuminated areas and on the spontaneous exploratory behaviour of the animals, applying mild stressors i.e. novel environment and light. The test apparatus consists of a small dark secure compartment (one third) and a large illuminated aversive compartment (two thirds). 2. The test was developed with male mice. The strain, weight and age may be crucial factors. 3. The extent to which an anxiolytic compound can facilitate the exploratory activity depends on the baseline level in the control group. Differences between the type and severity of external stressors might account for variable results reported by different laboratories. 4. In conclusion, the black and white test may be useful to predict anxiolytic-like or anxiogenic-like activity in mice. Transitions have been reported to be an index of activity-exploration because of habituation over time and the time spent in each compartment to be a reflection of aversion. Classic anxiolytics (benzodiazepines) as well as the newer anxiolytic-like compounds (e.g. serotonergic drugs) can be detected using this paradigm. It has the advantages of being quick and easy to use, without requiring the prior training of animals.

Animal models of anxiety in mice.

Among the multiple possibilities to study human pathologies, animal models remain one of the most used pathways. They allow to access to unavailable answers in human patients and to learn about mechanisms of action of drugs. Primarily developed with rats, animal models in anxiety have been adapted with a mixed success for mice, an easy‐to‐use mammal with better genetic possibilities than rats. In this review, we have focused on the most used animal models in anxiety in mice. Both conditioned and unconditioned models are described, to represent all types of animal models of anxiety. Behavioural studies require strong care for variable parameters, linked to environment, handling or paradigm; we have discussed about this topic. Finally, we focused on the consequences of re‐exposure to the apparatus. Test–retest procedures can bring in new answers, but should be deeply studied, to revalidate the whole paradigm as an animal model of anxiety.

Anxiolytic-like effects of 5-HT2 ligands on three mouse models of anxiety

The behavioural effects of 5-HT(2) receptor agonists, 5-HT(2A) and 5-HT(2C) receptor antagonists were investigated in the mouse four plates test (FPT), light/dark paradigm (L/D) and the elevated plus maze (EPM), in order to elucidate the role of the 5-HT(2) receptor subtypes in these models and to address the inconclusive results previously reported using rat psychopharmacological models. All compounds were administered intraperitoneally 30 min before each test. DOI, a preferential 5-HT(2A) agonist (0.5-8 mg/kg) and BW 723C86, a 5-HT(2B) agonist (8 and 16 mg/kg) provoked an anxiolytic-like response in the FPT. In the EPM, an anxiolytic-like effect was observed for DOI (0.5, 1 and 2 mg/kg), BW 723C86 (0.5, 4, 8 and 16 mg/kg), RO 60-0175 a 5-HT(2C) agonist (4 mg/kg) and the non-selective 5-HT(2) receptor agonist mCPP (0.25 mg/kg.). Ketanserin, a 5-HT(2A/2C) non-selective receptor antagonist (0.015 and 0.03 mg/kg), induced an anxiogenic-like effect in the L/D paradigm. The 5-HT(2C) antagonists (RS 10-2221, SDZ SER082 and SB 206553) were without effect in all three tests. These behavioural results are indicative of an anxiolytic-like action of 5-HT(2) receptor agonists, an anxiogenic-like effect of 5-HT(2A) receptor antagonism, whereas the blockade of 5-HT(2C) receptors are without effect in the mouse models studied.

Antidepressant-like effects in various mice strains in the tail suspension test.

Several studies have reported rodent strain differences in the response to antidepressants in animal models of depression. The aim of the present study was to investigate the potential contribution of genetic factors to antidepressant response in an animal model of depression: the tail suspension test (TST). For this study four mice strains (Swiss and NMRI, two outbred strains and DBA/2 and C57BL/6J Rj, two inbred strains) were submitted to the TST after acute administration of five antidepressants: the tricyclic antidepressants (TCAs) imipramine and desipramine, the selective serotonin (5-HT) reuptake inhibitors (SSRIs) paroxetine and citalopram and the dopamine reuptake inhibitor bupropion. The C57BL/6J Rj strain had a longer baseline immobility time in comparison to the other strains. All antidepressants studied in this work decreased immobility time in the Swiss and C57BL/6J Rj strains. However, the Swiss strain displayed greater sensitivity to citalopram (from 2mg/kg) and C57BL/6J Rj to paroxetine (from 0.5mg/kg). This latter presented a greater size-effect with citalopram than with other strains and reached more than 60% from 8mg/kg. Moreover the size-effect of desipramine, paroxetine and bupropion in Swiss mice was greater than in the other strains in the TST. The NMRI and DBA/2 mice only responded to 5-HT reuptake inhibitors, both selective (paroxetine, citalopram) or non-selective (imipramine). The NMRI strain was more sensitive to imipramine and presented a size-effect (43% at 8mg/kg) superior to those of other strains. DBA/2 strain was more sensitive to citalopram than paroxetine and imipramine. Our results suggest that response to an antidepressant treatment is under control of genetic factors and that the strain of mouse is an important parameter to consider.

A new approach to the light/dark test procedure in mice

The effect of the known anxiolytic agents diazepam and alprazolam and a putative anxiogenic agent, FG 7142, were assessed in a fully automated and computer-integrated two-compartment light/dark apparatus. In addition, psychostimulant drugs (amphetamine, adrafinil, amineptine, and caffeine) were tested to determinate the influence of increasing locomotor activity on the indices of anxiety. Some modifications, such as using a soiled apparatus, have been made from the initial model to reduce any neophobic response to the test situation. These results have been compared to results obtained after cleaning between trials. In addition, strain differences have been assessed by comparing the effect of Swiss mice with the C57Bl/6J strain. The role of each parameter as an index of anxiety is discussed. The time spent in the lit area and exploratory behaviors seemed to be the most reliable parameter for assessing anxiolytic-like activity. Diazepam and alprazolam were found to have an anxiolytic profile. FG 7142 did not demonstrate any intrinsic effect. Amphetamine was reported to be anxiogenic, and amineptine, adrafinil, and caffeine only had a psychostimulant profile. We conclude that the light/dark test may be useful for identifying putative anxiolytic and anxiogenic agents, but an additional test such as an open field or an actimeter test must be performed as a control with regard to the problem of sedation and change in exploration. The Swiss strain of mice has been found a suitable strain to be used in the test.

Monoamine metabolism changes following the mouse forced swimming test but not the tail suspension test.

Microdialysis, binding and behavioural studies have shown that the dopaminergic system plays a role in antidepressant treatment. It has been suggested that stress may provoke a modification in dopamine (DA) release in different brain areas and that the forced swimming test (FST), in its own accord as a stressor, may be responsible for this modification. Naive male Swiss mice, receiving saline solution, were used in two animal models of depression, the FST and the tail suspension test (TST). In order to understand the locomotor aspect of each test, groups of mice were studied for effects on locomotor activity. Following each test, mice were killed by cervical dislocation, brains were removed and concentrations of amines in the whole brain were analysed by high‐performance liquid chromatography. DA concentration increased from 5 min of the FST, dihydroxyphénylacetate (DOPAC), from 20 min of FST and serotonin, from 8 min of FST. No modification of noradrenaline was observed during the FST and no modification of the neurotransmitter concentrations was observed during the TST. Following an FST of 2‐min duration, a hypolocomotor effect was observed in the subsequent actimeter test. The same effect was observed after a TST of 8 min and onwards. This study confirms the fact that although these two tests are used to study depression, they involve different neuronal mechanisms.

Anxiolytic-like effects of antidepressants after acute administration in a four-plate test in mice.

The four-plate test (FPT) is an animal model of anxiety based on stress caused by unconditioned fear. An increase of spontaneous punished behavior was used as a measure to determine the anxiolytic effects of various antidepressants (ADs). In the present study. ADs with different mechanisms of action, including tricyclics, selective serotonin reuptake inhibitors (SSRIs), a monoamine oxidase inhibitor (MAOI), and atypicals, were studied in the FPT to evaluate their anxiolytic-like effects following acute administration. The number of punished crossings was dramatically increased by the SSRIs citalopram, fluvoxamine, and paroxetine, but not fluoxetine. The mixed 5-HT/NE reuptake inhibitors, milnacipran and venlafaxine, also demonstrated strong antipunishment effects. The specific NE reuptake inhibitors, desipramine and maprotiline, and the atypical AD trazodone, enhanced freezing behavior, suggesting anxiogenic-like behavior. It was concluded that, in the FPT, a model based on spontaneous response, where animals are exposed to an aversive environment from which they can only escape by being motionless, this kind of behavior might be related to anticipatory anxiety. In this situation, ADs acting preferentially on 5-HT transmission possessed clear anxiolytic like effects. The balance between the two transmitters, 5-HT and NE, seemed to be a crucial factor.

Additive effects of lithium and antidepressants in the forced swimming test: further evidence for involvement of the serotoninergic system

In the mouse forced swimming test (FST) pretreatment with a subactive dose of lithium (1 mEq/kg), given IP 45 min before the test, facilitated the antidepressant activity of iprindole, fluoxetine, and moclobemide (given IP 30 min before the test). These antidepressants (ADS) were not active alone in the FST in this study. Moreover, when subactive lithium was combined with a wide range of ADS, each given at subactive doses, those ADS with serotoninergic properties (e.g. imipramine, citalopram, paroxetine, fluoxetine, trazodone, mianserin, and moclobemide) significantly reduced immobility times. ADS acting primarily on noradrenaline (NA) or dopamine (DA) systems (desipramine, maprotiline, viloxazine, and bupropion) did not significantly decrease immobility when given in combination with lithium. This was also the case for RO 16 6491 [a reversible, B specific monoamine oxidase inhibitor (MAOI)], nialamide, and pargyline (both irreversible, mixed MAOIs). The anti-immobility effect of iprindole in combination with lithium suggests either a direct or indirect action on the serotonin (5HT) system by this ADS whose mechanism of action remains obscure. These results, using an animal behavioral model of depression and combining our present knowledge of the acute action of various ADS, support the hypothesis that the potentiation by lithium of ADS is via direct 5HT mechanisms,indirectly via a NA/5HT link, and/or by second messenger systems. Lithium may also facilitate the expression of antidepressant activity of ADS not active by themselves in the FST.

Evaluation of efficacies of different classes of antidepressants in the forced swimming test in mice at different ages

1. The efficacies of different classes of antidepressants were investigated using the forced swimming test with mice at different ages. 2. Imipramine (4-32 mg/kg), desipramine (2-16 mg/kg) and bupropion (32, 64 mg/kg) showed activity in all age groups. 3. The selective serotonin reuptake inhibitors (SSRIs) citalopram (16 and 32 mg) and paroxetine (4 and 8 mg) were inactive in the oldest (40 weeks) group of mice, despite showing activity at the same doses in mice ranging in age from 4-24 weeks old. 4. Both SSRIs showed anti-immobility effects at low doses, (paroxetine: 1 and 2 mg/kg; citalopram: 4 and 8 mg/kg) in the 40-week old mice. These effects were not evident in the three younger groups of mice. 5. Moclobemide, a reversible selective inhibitor of monoamine oxidase-A, showed activity only at a high dose (128 mg/kg) and only in 12-week old animals. 6. Since SSRIs have been reported to have relatively selective effects on 5-HT1B receptors, the present results suggest that further studies comparing the effectiveness of SSRIs and other antidepressants in elderly patients should be done. Studies of the effects of aging on the density and/or affinity of 5-HT1A and 5-HT1B/1D receptors are also warranted.