The social behavior of honeybees (Apis mellifera) has been extensively investigated, but little is known about its neuronal correlates. We developed a method that allowed us to record extracellularly from mushroom body extrinsic neurons (MB ENs) in a freely moving bee within a small but functioning mini colony of approximately 1,000 bees. This study aimed to correlate the neuronal activity of multimodal high-order MB ENs with social behavior in a close to natural setting. The behavior of all bees in the colony was video recorded. The behavior of the recorded animal was compared with other hive mates and no significant differences were found. Changes in the spike rate appeared before, during or after social interactions. The time window of the strongest effect on spike rate changes ranged from 1 s to 2 s before and after the interaction, depending on the individual animal and recorded neuron. The highest spike rates occurred when the experimental animal was situated close to a hive mate. The variance of the spike rates was analyzed as a proxy for high order multi-unit processing. Comparing randomly selected time windows with those in which the recorded animal performed social interactions showed a significantly increased spike rate variance during social interactions. The experimental set-up employed for this study offers a powerful opportunity to correlate neuronal activity with intrinsically motivated behavior of socially interacting animals. We conclude that the recorded MB ENs are potentially involved in initiating and controlling social interactions in honeybees.
