Individual differences in fear memory expression engage distinct functional brain networks

Abstract Fearful stimuli elicit a mix of active (e.g., evasion) and passive (e.g., freezing) behaviors in a wide range of species, including zebrafish (Danio rerio). However, it is not clear if individual differences in fear responses exist and, if so, what parts of the brain may underlie such differences. To probe these questions, we developed a contextual fear conditioning paradigm for zebrafish that uses conspecific alarm substance (CAS) as an unconditioned stimulus where fish associate CAS administration with a specific tank. To identify individual differences, we collected behavioral responses from over 300 fish from four different strains (AB, TU, TL, and WIK) and both sexes. We found that fear memory behavior fell into four distinct groups: non-reactive, evaders, evading freezers, and freezers. We also found that background strain and sex influenced how fish respond to CAS, with males more likely to increase evasive behaviors than females and the TU strain more likely to be non-reactive. Finally, we performed whole-brain activity mapping to identify the brain regions that are associated with different behavioral responses. All groups exposed to the tank had strong engagement of the telencephalon, whereas regions beyond the telencephalon distinguished behavioral groups: animals that have high levels of freezing, but low levels of evasion, uniquely engage the cerebellum, preglomerular nuclei, and pretectal areas, whereas those fish that mix evasion with freezing engage the preoptic and hypothalamic areas. Taken together, these findings reveal that zebrafish exhibit individual differences in fear memory expression that are supported at the neural level by extra-telencephalic regions.