# Endocytic protein AP180 assembly domain regulates synaptic vesicle size and release in Caenorhabditis elegans

## TL;DR

This study shows how the AP180 protein controls synaptic vesicle size and release in C. elegans through its Assembly Domain and actin binding.

## Contribution

The study reveals that the AP180 Assembly Domain regulates vesicle size and release via actin interactions, influencing neurotransmission.

## Key findings

- AP180's C-terminal Assembly Domain determines synaptic vesicle size and regulates release properties.
- Enlarged vesicles from AP180∆AD escape curvature-dependent inhibition by complexin but remain dependent on it for evoked responses.
- Replacing the AP180 AD with actin-binding motifs restores normal vesicle size and release frequency.

## Abstract

Neuronal communication relies on neurotransmitter release from synaptic vesicles. The endocytic protein AP180 is critical for efficient vesicle recycling at presynaptic terminals, and its loss impairs neurotransmission, producing reduced release frequency, enlarged synaptic vesicles, and increased quantal amplitude. Yet how AP180 controls vesicle size and whether vesicle size influences release remains unclear. Here, we show that the C-terminal Assembly domain (AD) of AP180 determines vesicle size and thereby regulates release properties in Caenorhabditis elegans. An AP180 variant lacking the AD (AP180∆AD) increases release frequency, contrasting sharply with the reduced transmission in ap180 null mutants, yet fails to correct the vesicle size or quantal amplitude. These enlarged vesicles evade curvature-dependent inhibition by complexin, a presynaptic regulator of fusion, while remaining dependent on complexin for evoked responses. This selective escape reveals that vesicle size influences release dynamics through curvature-sensing proteins. Replacing the AP180 AD with actin-binding motifs restores normal vesicle size, quantal amplitude, and release frequency, indicating that actin interactions are both necessary and sufficient for AD function. Biochemically, we show that the intrinsically disordered AD forms condensates that enrich actin monomers and nucleate filament assembly, while full-length AP180 couples PIP2-rich membranes to actin filaments. Together, these findings reveal that the AP180 AD regulates synaptic vesicle size through actin binding, establishing vesicle morphology as a key influencer of curvature-dependent release control.

The endocytic protein AP180 regulates synaptic vesicle recycling during neurotransmission, but how vesicle size and release frequency are determined remains unclear. This study shows that AP180 Assembly Domain regulates synaptic vesicle size and release fidelity in C. elegans through actin binding, revealing how vesicle morphology influences neurotransmission.