Selective genetic targeting of the mouse efferent vestibular nucleus identifies monosynaptic inputs and indicates...

The vestibular system is a critical sensory modality required for coordinated movement, balance, and our ability to interact with the surrounding environment. Vestibular sensory neurons provide the nervous system with information about head rotation and acceleration. However, the nervous system can also modify the activity of sensory neurons and hair cells via the actions of the efferent vestibular system (EVS). The function of the EVS has remained unknown partly because of an inability to target efferent vestibular neurons in a selective manner to understand their synaptic inputs and function during behavior. Here, we present a novel method for the selective targeting and expression of flp-recombinase in EVS neurons. We take advantage of the dual expression of choline acetyl transferase (ChAT) and calcitonin gene-related peptide (CGRP) in these neurons to develop an adeno-associated virus (AAV) that expresses a gene only in neurons with this intersectional expression. We use this system to map the monosynaptic inputs to EVS neurons and show inputs from distinct populations of brainstem and midbrain regions, indicating a functional role as a multimodal processing center and integrator for the vestibular periphery. To demonstrate the applicability of our technology in behavioral assays, we performed a preliminary behavior analysis (treadmill running and open field) in mice with disrupted EVS function. Although more bespoke assays are required to ascertain EVS function(s), our viral method presents a novel tool for investigators examining the role of the vestibular system and its central circuits.NEW & NOTEWORTHY We provide a novel means of targeting and accessing the EVN and provide valuable new information on the anatomical organization of the efferent vestibular system. For the first time, we present a holistic understanding of the type of information that the EVN receives and processes, highlighting the multimodal nature of the EVN, acting as a hub to update the vestibular periphery in real time with relevant internal and external information to support overall vestibular health.