Table 1.
An overview of social preference test in zebrafish.
| Assessed variable | Strain | Tested fish per group | Sex of tested fish | Age of tested fish | Test tank characteristics | Social stimulus | Behavioral protocols | Results | References |
|---|---|---|---|---|---|---|---|---|---|
| STUDIES CARRIED OUT IN UNTREATED WT ZEBRAFISH | |||||||||
| Phenotype and rearing management | AB and nacre | 33–38 | Mixed | 5–6 months | One 245 L tank (122 × 32 × 55 cm, L × W × H) divided into three chambers: one for AB, one for nacre and a central one for the tested fish | 4 AB vs. 4 nacre fish | Habituation phase: 10 min; interaction phase: 15 min | Zebrafish showed positive social preference with individuals of the same phenotype with which they were raised | (51) |
| Visual characteristics of animated zebrafish images | SF | 10 | 50-50% m-f | 6–8 months | One 40 L tank (51 × 30 × 25 cm, L × W × H) with flat LCD computer screens for social stimuli on its left and right sides | 1 modified vs. 1 unmodified computer-animated female zebrafish | Habituation phase: 10 min; interaction phase: 5 min | Visual characteristics of animated zebrafish images affected social preference | (19) |
| Body size of conspecifics | n.a. | 6–7 | Males | 3–12 months | One 14 L tank (58 × 15 × 15 cm, L × W × H) divided into three chambers: one for large conspecifics, one for small conspecifics, and a central one for tested fish | 3 large vs. 3 small conspecifics | Habituation phase: 10 min; interaction phase: 10 min | Zebrafish males preferred to socialize with larger body size conspecifics compared with smaller ones | (49) |
| Familiarity of conspecifics | AB | n.a. | n.a. | n.a. | One tank (19 × 13.2 × 9.3 cm, L × W × H) divided into five chambers: one for familiar conspecifics, one for unfamiliar conspecifics, two empty, and a central one for tested fish | 3 familiar vs. 3 unfamiliar conspecifics | Habituation phase: n.a.; interaction phase: 5 min | Zebrafish showed increased social preference for unfamiliar over familiar conspecifics | (26) |
| Temperature increase from 26 to 34°C | AB | 15 | 50-50% m-f | 12 months | Three aligned 4 L tanks (25 × 28 × 16 cm, L × W × H). One for the social stimulus, one empty for no social stimuli, and a central one for tested fish | 3 conspecifics | Habituation phase: n.a.; interaction phase: 10 min | Heat treatment reduced social preference | (52) |
| STUDIES CARRIED OUT IN PHARMACOLOGICALLY TREATED WT ZEBRAFISH | |||||||||
| Alcohol treatment, chronic (10 days in 0.50% alcohol v/v) and then acute (1 h in 0.25, 0.50, or 1.00% alcohol v/v) | AB and SF | 13–18 | 50-50% m-f | 12 months | One 37 L tank (50 × 25 × 30 cm, L × W × H) with flat LCD computer screens, for displaying social stimuli, on its left and right sides | 5 computer-animated conspecifics | Habituation phase: 8 min; interaction phase: 8 min | AB fish exposed to chronic and subsequently to 1.00% acute alcohol treatment showed reduced social preference, while SF fish did not | (53) |
| Lysergic acid diethylamide (LSD) treatment (20 min in 5, 25, 50, 75, 100 or 250 μg/L) | SF | 10 | Mixed | 5–7 months | Five areas of a modified T-maze (50 × 10 × 10 cm, L × W × H). One chamber for the social stimulus, one empty chamber for no social stimuli, and the central chamber (divided into three areas), for tested fish | 1 conspecific | Habituation phase: 30 s; interaction phase: 6 min | LSD treatment did not influence zebrafish social preference | (54) |
| Ketamine treatment (20 min in 2, 20, or 40 mg/L) | SF | 12 | Mixed | 5–7 months | Five areas of a modified T-maze (50 × 10 × 10 cm, L × W × H). One chamber for the social stimulus, one empty for no social stimuli, and the central chamber (divided into three areas), for tested fish | 1 conspecific | Habituation phase: 30 s; interaction phase: 6 min | Ketamine treatment did not affect zebrafish social preference | (55) |
| MK-801 treatment (30 min in 2, 20, or 100 μM) | SF | 8 | 50-50% m-f | 6–8 months | Three aligned 2 L tanks (21 × 10 × 10 cm, L × W × H). One for social stimulus, one empty for no social stimuli, and a central one for tested fish | 5 conspecifics | Habituation phase: n.a.; interaction phase: 30 min | MK-801, at the highest dose (100 μM), significantly reduced fish social preference | (56) |
| Isotocin and vasotocin injection (0.001–40 ng/kg body weight) | n.a. | 10 | 50-50% m-f | 6–12 months | One tank (122 × 32 × 55 cm, L × W × H) divided into three chambers: one for WT conspecifics, one for nacre mutants, and a central one for tested fish | 4 conspecifics | Habituation phase: 5 min; interaction phase: 15 min | Both neuropeptides increased social preference in a dose-dependent manner | (57) |
| D1-receptor antagonist SCH23390 treatment (30 min in 0.1 or 1.0 mg/L) | AB and SF | 20 | 50-50% m-f | 4–6 months | One 37 L tank (50 × 25 × 30 cm, L × W × H) with flat LCD computer screens for social stimuli on its left and right sides | 5 computer-animated female zebrafish conspecifics | Habituation phase: 8 min; interaction phase: 8 min | In AB fish treated with the high concentration of SCH23390, social preference was impaired. In the SF strain, no significant effects were observed | (27) |
| Indole alkaloid ibogaine treatment (20 min 10 or 20 mg/L) | SF | 15 | 50-50% m-f | 5–8 months | Five chambers of a modified T-maze (50 × 10 × 10 cm, L × W × H). One chamber for social stimulus, one empty for no social stimuli, and the central three chambers for tested fish | 1 conspecific | Habituation phase: 30 s; interaction phase: 6 min | Ibogaine treatment did not influence zebrafish social preference | (58) |
| Injection of isotocin, an isotocin antagonist, or vasotocin, a vasotocin antagonist (10 μg/g body weight) | n.a. | 20 | 100% females | 4–5 months | One tank (150 × 50 cm, L × W) divided into three chambers: one for social stimulus, one empty for no social stimuli, and a central one for tested fish | 8 conspecifics | Habituation phase: n.a.; interaction phase: 10 min | Vasotocin and its antagonist decreased social preference, while isotocin and its antagonist had no significant effects on social preference | (59) |
| MK-801 treatment (1 h in 100 μM) or acute alcohol treatment (1 h in 0.125 or 0.50% alcohol v/v) | AB | 177 for MK-801, 96 for low and 82 for high alcohol treatments | n.a. | 3 weeks | One tank (4 × 3.2 cm, L × W) divided into three chambers: one for social stimulus, one empty for no social stimuli and a central one for tested fish | 1 or 3 conspecifics | Habituation phase: 15 min; interaction phase: 15 min | After blocking of NMDA receptors, fish exhibited no social preference. Alcohol treatment significantly reduced social preference only at the higher dose | (60) |
| Injection of amphetamine derivatives DOB (0.05–2 mg/kg), PMA (0.0005–2 mg/kg), MDMA (0.25–20 mg/kg) or ritanserin (0.025–2.5 mg/kg body weight) in association with the maximal doses of DOB, PMA or MDMA | SF | 10 | 50-50% m-f | 6–12 months | One tank (122 × 32 × 55 cm, L × W × H) divided into five areas: the outermost chambers as stimulus areas with pictures of zebrafish as social stimuli, and the central three areas (1 chamber) for tested fish | Pictures of 6 conspecifics (SF vs. nacre) | Habituation phase: 5 min; interaction phase: 15 min | Inverted-U shape dose-dependent increase in social preference was observed for DOB, PMA and MDMA treatments | (61) |
| Fluoxetine (15 min in 50 μg/L) and diazepam (15 min in 16 μg/L) | SF | 7–10 | 50-50% m-f | 6 months | Three aligned tanks (30 × 15 × 10 cm, L × W × H). One for social stimulus, one empty for no social stimuli, and a central one for tested fish | 15 conspecifics | Habituation phase: 30 s; interaction phase: 10 s | Fluoxetine (15 min in 50 μg/L) and diazepam (15 min in 16 μg/L) decreased social preference | (62) |
| MK-801 treatment (15 min in 5 μM) and subsequent oxytocin, carbetocin or L-368,899 injection (10 ng/kg body weight) | SF (Tübingen) | 16 | n.a. | 6–8 months | Three aligned tanks (30 × 10 × 15 cm, L × W × H). One for social stimulus, one empty for no social stimuli, and a central one for tested fish | 15 conspecifics | Habituation phase: 30 s; interaction phase: 5 min | MK-801 induced a decrease in social preference. Oxytocin and carbetocin re-established this behavior, while L-368,899 did not | (63) |
| Sodium valproate treatment, chronic (from 1 dpf 7 h per day for 6 days in 20 μM) or acute (at 1 dpf 7 h in 100 μM) | AB | 60 | n.a. | 1 month | One tank divided into two chambers: one for social stimulus and one for tested fish (7 × 4.2 cm, L × W). | 6 conspecifics | Habituation phase: 20 min; interaction phase: 30 min | Chronic exposure to sodium valproate significantly impaired social preference, while acute exposure did not | (45) |
| Acute alcohol treatment (at 1dpf 2 h in 0.01, 0.25, 0.50, or 1.00% alcohol v/v) | SF | 8 | 50-50% m-f | 4 months | Three aligned tanks. One for social stimulus, one empty for no social stimuli, and a central one for tested fish. | 3 conspecifics | Habituation phase: n.a.; interaction phase: 6 min. | Embryonal exposure to alcohol reduced social preference in a dose dependent manner | (64) |
| Acute alcohol treatment in association with taurine treatment (1 h in 0.25% alcohol v/v and in 42, 150 and 400 mg/ L of taurine) | SF | 10–12 | 50-50% m-f | 4–6 months | One tank (25 × 10 × 15 cm, L × W × H) divided into three chambers: one for social stimulus, one empty for no social stimuli, and a central one for tested fish | 4 conspecifics | Habituation phase: 30 s; interaction phase: 1 min | Acute alcohol exposure in association with the highest dose of taurine significantly reduced social preference | (21) |
| Formalin-inactivated Aeromonoas hydrophila bacterin injection (50 μL of 4 × 105) | n.a. | 12 | Mixed | 6 months | One tank (24 × 8 × 20 cm, L × W × H) divided into three chambers: one for social stimulus, one empty for no social stimuli, and a central one for tested fish | 15 conspecifics | Habituation phase: n.a.; interaction phase: 1 min | Formalin-inactivated Aeromonoas hydrophila bacterin injection reduced social preference | (17) |
| Untreated tannery effluent exposure (30 days in 0.1 and 0.3% of body biomass) | n.a. | 15 in each group | Mixed | 6–8 months | Three aligned 5 L tanks (20 × 15 × 20 cm, L × W × H). One for social stimulus, one empty for no social stimuli, and a central one for tested fish | 6 conspecifics | Habituation phase: 4 min; interaction phase: 4 min | Untreated tannery effluent at the highest dose reduced social preference | (20) |
| Alcohol treatment, intermittent acute (20 min per day for 4 days in 1% alcohol v/v) and intermittent chronic (20 min per day for 16 days in 1% alcohol v/v) | SF | 25 in each group | Mixed | 4–5 months | Three aligned tanks. One for social stimulus, one empty for no social stimuli, and a central one for tested fish | 5 conspecifics | Habituation phase: 2 min; interaction phase: 5 min | Acute alcohol exposure significantly reduced social preference, while chronic exposure did not | (65) |
| Chronic acesulfame treatment (2 months in 1, 10 or 100 mg/L) | AB | 12 in each group | n.a. | 9 months | One tank (30 × 10 × 10 cm, L × W × H) divided into three chambers: one for social stimulus and two for tested fish | 6 conspecifics | Habituation phase: n.a.; interaction phase: 3 min | Chronic acesulfame exposure reduced social preference | (66) |
| L-368,899 injection (100 μg/g body weight). | AB | 21 in each group | 50-50% m-f | n.a. | One tank (90 × 30 cm, L × W) divided into three chambers: one for social stimulus, one empty for no social stimuli, and a central one for tested fish | 8 conspecifics | Habituation phase: 10 min; interaction phase: 15 min | L-368,899 injection reduced social preference | (50) |
| L-368,899 treatment (1 h in 100 μM) | AB | 77–87 in each group | n.a. | 3 weeks | One tank (4 × 3.2 cm, L × W) divided into three chambers: one for social stimulus, one empty for no social stimuli and a central one for tested fish | 1 conspecific | Habituation phase: 15 min; interaction phase: 15 min | L-368,899 treatment reduced social preference | (50) |
| STUDIES CARRIED OUT IN GENETICALLY ENGINEERED ZEBRAFISH | |||||||||
| otpa and otpb | 10 in each group | n.a. | n.a. | One tank divided into three chambers: one for social stimulus, one empty for no social stimuli, and one for tested fish | 4 conspecifics | Habituation phase: 5 min; interaction phase: 9 min | otpa mutants showed reduced social preference as compared with WT and otpb mutants | (67) | |
| reln | 10–12 in each group | Mixed | 3–6 months | One tank divided into five chambers: one for social stimulus, three empty, and a central one for tested fish | 3 conspecifics | Habituation phase: n.a.; interaction phase: 5 min | reln mutants showed similar social preference as compared with WT | (68) | |
| dyrk1aa | 6–7 in each group | Males | 3–12 months | One 14 L tank (58 × 15 × 15 cm, L × W × H) divided into three chambers: one for large conspecifics, one for small conspecifics, and a central one for tested fish | 3 large vs. 3 small WT conspecifics | Habituation phase: 10 min; interaction phase: 10 min | dyrk1aa heterozygous zebrafish exhibited a significantly higher preference for the large conspecifics, while homozygous KO zebrafish showed no such preference | (49) | |
| oxtr | 10–14 in each group | Mixed | 3–6 months | One tank (20 × 19 × 5 cm, L × W × H) divided into three chambers: one for social stimulus, one empty for no social stimuli, and one for tested fish | 4 conspecifics | Habituation phase: 5 min; interaction phase: 10 min | oxtr mutants showed normal social preference | (69) | |
n.a, data not available; m-f, males-females ratio; SF, short fin; DOB, 2,5-dimetoxy-4-bromo-amphetamine hydrobromide; PMA, para-methoxyamphetamine; MDMA, 3,4 methylene-dioxymethamphetamine; dpf, days post-fertilization.