Searching for learning-dependent changes in the antennal lobe: simultaneous recording of neural activity and aversive olfactory learning in honeybees

Plasticity in the honeybee brain has been studied using the appetitive olfactory conditioning of the proboscis extension reflex, in which a bee learns the association between an odor and a sucrose reward. In this framework, coupling behavioral measurements of proboscis extension and invasive recordings of neural activity has been difficult because proboscis movements usually introduce brain movements that affect physiological preparations. Here we took advantage of a new conditioning protocol, the aversive olfactory conditioning of the sting extension reflex, which does not generate this problem. We achieved the first simultaneous recordings of conditioned sting extension responses and calcium imaging of antennal lobe activity, thus revealing on-line processing of olfactory information during conditioning trials. Based on behavioral output we distinguished learners and non-learners and analyzed possible learning-dependent changes in antennal lobe activity. We did not find differences between glomerular responses to the CS+ and the CS− in learners. Unexpectedly, we found that during conditioning trials non-learners exhibited a progressive decrease in physiological responses to odors, irrespective of their valence. This effect could neither be attributed to a fitness problem nor to abnormal dye bleaching. We discuss the absence of learning-induced changes in the antennal lobe of learners and the decrease in calcium responses found in non-learners. Further studies will have to extend the search for functional plasticity related to aversive learning to other brain areas and to look on a broader range of temporal scales.