# Two types of axonal muscarinic acetylcholine receptors mediate formation of saliva cocktail in the tick Ixodes ricinus

## TL;DR

The study identifies two types of acetylcholine receptors in ticks that control saliva production, offering new ways to disrupt tick feeding and pathogen transmission.

## Contribution

Discovery of two distinct muscarinic acetylcholine receptor types in ticks and their role in saliva regulation.

## Key findings

- Two pharmacologically distinct mAChRs (type A and B) were identified in Ixodes ricinus.
- Type B mAChRs show atypical signaling, suggesting unconventional receptor function.
- mAChR types coordinate fluid and protein secretion via distinct salivary gland regions.

## Abstract

Hard ticks depend upon an ability to precisely and dynamically regulate their saliva to successfully evade host haemostatic and immune defences during extended blood feeding. Although pilocarpine, an exogenous muscarinic acetylcholine receptor (mAChR) agonist, can stimulate salivation experimentally, the endogenous control of saliva secretion by acetylcholine remains poorly understood. Here, we identify and characterise two pharmacologically distinct mAChRs (type A and B) in the genome of the medically important tick Ixodes ricinus. Molecular dynamics simulations and targeted mutagenesis reveal that type B mAChRs exhibit an atypical muscarinic profile, suggesting unconventional receptor signalling. Combining immunolabelling, in vivo pharmacology, and proteomics, we show that specific central neurons interact with distinct salivary gland regions via mAChR type-specific axons, coordinating fluid and protein secretion through separate acini and likely acting upstream of a neuropeptide-dependent cascade. This previously unrecognised mechanism of neural control offers new insights into how ticks modulate their saliva advancing our understanding of vector-host interactions, with potential implications for disrupting pathogen transmission.

Here authors demonstrate that distinct muscarinic receptor types drive coordinated salivary output in the tick Ixodes ricinus. Mapping the underlying circuits reveals functional neural pathways that could be targeted to disrupt tick feeding.