# Integrated Transcriptomic and Metabolomic Insights into “I See You” (ISY) Defensive Behavior in Apis cerana Against Vespa velutina

## TL;DR

This study explores how honeybees defend against hornets by analyzing brain gene and molecule changes during a defensive behavior called 'I See You'.

## Contribution

The study integrates transcriptomic and metabolomic data to reveal molecular mechanisms behind a honeybee's defensive response to hornet attacks.

## Key findings

- ISY behavior is linked to changes in brain messengers like serotonin and dopamine.
- Energy-supplying molecules increase during ISY behavior.
- Purine and tyrosine metabolism pathways are upregulated in response to hornet attacks.

## Abstract

Hornets are major predators of honey bees. When a hornet approaches the nest, guard bees of Apis cerana perform an “I See You” (ISY) display. To explore what supports this rapid, coordinated display, we measured changes in brain gene activity and small molecules in bees performing ISY versus routine activity. ISY was associated with shifts in key brain messengers (including serotonin and dopamine) and with increases in molecules that supply energy. Together, these changes may help bees send signals faster and act in sync, sustaining the defensive wave. Understanding the biology behind shimmering offers a simple explanation for how colonies deter hornets and may provide practical clues.

Vespa velutina (V. velutina) is a globally widespread predator of honeybees, posing a serious threat to them. Apis cerana (A. cerana) has acquired the ability to partially thwart the predation of the wasps, but the molecular mechanism by which it responds to V. velutina attacks is unknown. Here, both transcriptomic and metabolomic analyses were integrated to decipher what happened in the brain tissues of A. cerana exposed to V. velutina attacks. Transcriptomic analysis revealed 296 differentially expressed genes (DEGs) that were significantly enriched in neural signaling pathways (particularly serotonin and dopamine transmission), the cAMP signaling pathway, and energy metabolism. Metabolomic results showed that V. velutina attacks affected 38 metabolic pathways involving 86 differentially expressed metabolites (DEMs), primarily including amino acids, nucleotides, and carbohydrates. Several metabolites, such as isocitrate, xanthine, and adenosine, were upregulated in response to the hornet presence. Combined pathway analysis indicated an upregulation of the purine and tyrosine metabolism pathways, highlighting their critical role in A. cerana against invasive hornets. The results of this study will help to understand the molecular mechanisms of A. cerana in response to V. velutina attacks and provide valuable insights for enhancing collective defensive behavior against V. velutina in honeybees.