Stimulus intensity and temporal configuration interact during bimodal learning and memory in honey bees

Multimodal integration is a core neural process with a keen relevance during ecological tasks requiring learning and memory, such as foraging. The benefits of learning multimodal signals imply solving whether the components come from a single event. This challenge presumably depends on the timing and intensity of the stimuli. Here, we used simultaneous and alternate presentations of olfactory and visual stimuli, at low and high intensities, to understand how temporal and intensity variations affect the learning of a bimodal stimulus and its components. We relied on the conditioning of the proboscis extension response (PER) to train honey bees to an appetitive learning task with bimodal stimuli precisely controlled. We trained bees to stimuli with different synchronicity and intensity levels. We found that synchronicity, order of presentation, and intensity significantly impacted the probability of exhibiting conditioned PER responses and the latency of the conditioned responses. At low intensities, synchronous bimodal inputs produced maximal multisensory enhancement, while asynchronous temporal orders led to lower performances. At high intensities, the relative advantage of the synchronous stimulation diminished, and asynchronous stimuli produced similar performances. Memory retention was higher for the olfactory component and bimodal stimuli compared to the visual component, irrespective of the training's temporal configuration. Bees retained the asynchronous bimodal configuration to a lesser extent than the synchronous one, depending on the stimulus intensity. We conclude that time (synchrony), order of presentation, and intensity have interdependent effects on bee learning and memory performance. This suggests caution when assessing the independent effects of each factor.