C57BL/6J mice fail to exhibit preference for social novelty in the three-chamber apparatus

Abstract

Laboratory models of neurodevelopmental disorders may be useful in assessing investigation and preference for social partners in mice. One such mouse model, the three-chamber test, is increasingly used as an index of social preference. The first phase measures preference for a social stimulus over an identical chamber without a stimulus mouse. The second phase measures preference for a novel mouse compared to the familiar mouse when the latter is presented in the previously empty chamber. In this study we provided an additional analysis of the second phase of the three-chamber test procedure, reversing the typical placement of the novel and familiar stimulus animals. In the first study, male C57BL/6J mice subjects encountered C57BL/6J stimuli and preferred a novel mouse over an empty chamber but failed to show a preference for the novel mouse in Phase 2 when the stimuli presentation was reversed. In an additional study, male C57BL/6J subjects encountered outbred CD-1 mice as stimuli, showing no significant novelty preference in either phase. Specific behavioral indices of investigation were similar to these duration findings with no enhancement of investigation when the novel stimulus mouse was encountered in the chamber in which the initial social stimulus was presented. These data suggest that C57BL/6J mice may show enhanced investigation/preference of novel social stimuli in the three-chamber test only when these stimuli are presented in a relatively novel context.

Fig 1

Three-chamber social approach and social novelty arena. Three interconnected chambers are separated by two manually operated sliding doors. Outside compartments contain inverted wire cups to house a stimulus mouse. A steel weight and a clear Plexiglas cylinder are placed on top of the inverted cups to prevent lifting or climbing on top. The inset shows a front view demonstrating the clear Plexiglas window on the front of the arena which permits videotaping of the dyadic interactions of the mice.

Fig. 2

Phase one sociability preference for a stimulus mouse of an unfamiliar B6 mouse (a) or for an unfamiliar CD-1 mouse (b) over an empty wire cup. * p<0.05, ** p<0.005, n=20/group. Data are expressed as Mean ± 1 S.E.M.

Fig 3

Increased frequency of contact (a) and increased frequency and duration of sniffing an unfamiliar B6 mouse (a,b) compared to the empty cup control. Increased frequency (c) and duration (d) of sniffing within the compartment with an unfamiliar CD-1 mouse compared to the empty cup control. White bars indicate the empty cup and black bars are the unfamiliar mouse stimulus. * p<0.05, ** p<0.001, n=20/group. Data are expressed as Mean ± 1 S.E.M.

Fig. 4

Mice spent significantly more time in the chamber associated with the novel B6 mouse over the familiar one but only in the standard condition (a). When CD-1 mice were used as stimuli, there was a significant increase in the duration spent in the chamber associated with the familiar mouse under the reversal condition (b). White bars represent familiar while black bars are novel stimuli. * p<0.05, ** p<0.01, n=10/group. Data are expressed as Mean ± 1 S.E.M.

Fig. 5

Behavioral data collected during phase 2 indicated a significant main effect for stimulus condition for sniffing frequency for B6 stimuli and post-hoc analyses revealed that under the standard presentation, mice spent more time sniffing the novel B6 mouse (a). * p<0.05, n=10/group. Data are expressed as Mean ± 1 S.E.M.