Who is the boss? Individual recognition memory and social hierarchy formation in crayfish

Under laboratory conditions, crayfish establish hierarchical orders through agonistic encounters whose outcome defines the dominant one and one, or more, submissive animals. These agonistic encounters are ritualistic, based on threats, pushes, attacks, grabs, and avoidance behaviors that include retreats and escape responses. Agonistic behavior in a triad of unfamiliar, size-matched animals is intense on the first day of social interaction and the intensity fades on daily repetitions. The dominant animal keeps its status for long periods, and the submissive ones seem to remember 'who the boss is'. It has been assumed that animals remember and recognize their hierarchical status by urine signals, but the putative substance mediating this recognition has not been reported. The aim of this work was to characterize this hierarchical recognition memory. Triads of unfamiliar crayfish (male animals, size and weight-matched) were faced during standardized agonistic protocols for five consecutive days to analyze memory acquisition dynamics (Experiment 1). In Experiment 2, dominant crayfish were shifted among triads to disclose whether hierarchy depended upon individual recognition memory or recognition of status. The maintenance of the hierarchical structure without behavioral reinforcement was assessed by immobilizing the dominant animal during eleven daily agonistic encounters, and considering any shift in the dominance order (Experiment 3). Standard amnesic treatments (anisomycin, scopolamine or cold-anesthesia) were given to all members of the triads immediately after the first interaction session to prevent individual recognition memory consolidation and evaluate its effect on the hierarchical order (Experiment 4). Acquisition of hierarchical recognition occurs at the first agonistic encounter and agonistic behavior gradually diminishes in the following days; animals keep their hierarchical order despite the inability of the dominant crayfish to attack the submissive ones. Finally, blocking of protein synthesis or muscarinic receptors and cold anesthesia impair memory consolidation. These findings suggest that agonistic encounters induces the acquisition of a robust and lasting social recognition memory in crayfish.