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Whole-brain monosynaptic inputs and outputs of glutamatergic
RV and AAV-helper were used for retrograde tracing process of the VN glutamatergic neurons. (From BrainVTA)
The viruses used in this article from BrainVTA are in the table below
Tracing Helper  PT-0023 rAAV2/9-Ef1α-DIO-RVG-WPRE-pA
 PT-0021 rAAV2/9-Ef1α-DIO-His-EGFP-2a-TVA-WPRE-pA
RV  RV-ENVA-△G-dsRed
Xunbei Shi, Haohua Wei, Zeka Chen, Jing Wang, Weimin Qu, Zhili Huang, Chunfu Dai
Pub Date: 2020-12-27, DOI: 10.1016/j.heares.2020.108159, Email: [email protected]
Vestibular nuclei complex (VN) glutamatergic neurons play a critical role in the multisensory and multimodal processing. The dysfunction of VN leads to a series of vestibular concurrent symptoms, such as disequilibrium, spatial disorientation, autonomic disorders and even emotion disorders. However, the reciprocal neural connectivity in the whole brain of VN glutamatergic neurons was incompletely understood. Here, we employed a cell-type-specific, cre-dependent, modified virus vector to retrogradely and anterogradely trace VN glutamatergic neurons in the VGLUT2-IRES-Cre mouse line. We identified and analyzed statistically the afferents and efferents of VN glutamatergic neurons in the whole brain, and also reconstructed monosynaptic inputs distribution of VN glutamatergic neurons at the three-dimensional level with the combination of a fluorescence micro-optical sectioning tomography system (fMOST). We found that VN glutamatergic neurons primarily received afferents from 57 nuclei and send efferents to 59 nuclei in the whole brain, intensively located in the brainstem and cerebellum. Projections from nuclei in the cerebellum targeting VN glutamatergic neurons mainly performed the balance control - the principal function of the vestibular system. In addition, VN glutamatergic neurons sent projections to oculomotor nucleus, trochlear nucleus and abducens nucleus dominating the eye movement. Except for the maintenance of balance, VN glutamatergic neurons were also directly connected with other functional regions, such as sleep-wake state (locus coeruleus, dorsal raphe nucleus, and laterodorsal tegmental nucleus, gigantocellular reticular nucleus, lateral paragigantocellular nucleus, periaqueductal gray, subcoeruleus nucleus, parvicellular reticular nucleus, paramedian raphe nucleus), and emotional regulation (locus coeruleus and dorsal raphe nucleus). Hence, this study revealed a comprehensive whole-brain neural connectivity of VN glutamatergic neurons and provided with a neuroanatomic foundation to further study on central vestibular circuits.

Figure 1. Retrograde tracing on VN glutamatergic neurons with a rabies virus-based, retrograde tracing system.
In this study, the authors employed the cell-type-specific retrograde and anterograde tracing vector to identify whole-brain long-range inputs and outputs of the glutamatergic neurons in the VN and served as a neuroanatomic basis to further research on vestibular circuits involving multisensory functions.
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