Territorial behaviors comprise a set of coordinated actions and response patterns found across animal species that promote the exclusive access to resources. House mice are highly territorial with a subset of males consistently attacking and chasing competing males to expel them from their territories and performing urine marking behaviors to signal the extent of their territories. Natural variation in territorial behaviors within a mouse colony leads to the formation of dominance hierarchies in which subordinate males can reside within the territory of a dominant male. While the full repertoire of such territorial behaviors and hierarchies has been extensively studied in wild-derived mice in semi-natural enclosures, so far they have not been established in the smaller enclosures and with the genetically-defined laboratory strains required for the application of neural recording and manipulation methods. Here, we present a protocol to rapidly induce an extensive repertoire of territorial behaviors in pairs of laboratory mice in an enclosure compatible with tethered neurocircuit techniques, including a method for the simultaneous tracking of urine marking behavior in mouse pairs. Using this protocol we describe the emergence of robust dominant-subordinate hierarchies between pairs of CD1 outbred or CD1xB6 F1 hybrid mice, but unexpectedly not in C57BL/6 inbred animals. Our behavioral paradigm opens the door for neurocircuit studies of territorial behaviors and social hierarchy in the laboratory.
© 2024. The Author(s).