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Updated: May 27, 2026
Assessing Social Dominance in Mouse Models Using the Tube Test
Published on: June 6, 2025
Assessing social rank in male mice using the multi-round warm spot test
Xiao Jia1, Can Tang1, Zhiyang Yu1
1Jiangsu Province Key Laboratory of Anesthesiology, Jiangsu Province Key Laboratory of Anesthesiology and Brain Science, School of Anesthesiology, Xuzhou Medical University, Xuzhou, Jiangsu, 221004, China.
Background:
The warm spot test exploits thermal preference to assess social dominance in mice, but traditional single-session protocols identify only the most dominant individual, leaving subordinate ranks unresolved.
Methods:
We developed a Multi-Round Warm Spot Test with stepwise elimination across four successive 10-minute rounds. Male C57BL/6 J mice (four per cage, n = 8 groups) were habituated and trained over two days. On test day, mice competed for a warm spot (34 °C) in a cold arena (0-4 °C). The mouse with greatest occupancy was removed after each round (Ranks 1-3). The final mouse remained alone in Round 4 as internal control (Rank 4). Independent validation was performed using the tube test.
Results:
Traditional testing identified only dominant individuals, with subordinates showing overlapping occupancy. The multi-round protocol successfully resolved all four ranks across all cages. Rank 1 mice dominated the first round, Rank 2 emerged after Rank 1 removal, and Rank 3 distinguished from Rank 4 in dyadic competition. Rank 4 mice showed significantly increased occupancy when alone (Round 4) versus dyadic competition (Round 3), recovering to training levels, confirming social suppression rather than differential thermal motivation. Tube test validation showed strong agreement with warm spot-derived ranks.
Conclusions:
The Multi-Round Warm Spot Test provides rapid, reliable complete hierarchy assessment in a single session, offering an efficient tool for studying social dominance mechanisms.
