Journal: eNeuro

Article Title: Disynaptic Inhibitory Cerebellar Control Over Caudal Medial Accessory Olive

doi: 10.1523/ENEURO.0262-23.2023

Figure Lengend Snippet: Anterograde transsynaptic tracing from MCN and LCN to IO via SC. A , Schematic representation of anterograde transsynaptic tracing strategy. B , Transsynaptic AAV1 expressing Cre and eGFP (cyan) was injected in the MCN ( n = 2) or LCN ( n = 2), followed by an injection of Cre-dependent mCherry (magenta) in the lateral part of SC (all animals). Bi – iv , Representative example photos of injection spots in MCN ( Bi ) and LCN ( Bii ) and subsequent transsynaptic labeling of cells in SC following injection in MCN ( Biii ) and LCN ( Biv ). Note that a small leakage to the interposed nucleus (INT) cannot be excluded in Bi and Bii , but the MCN and LCN injections never contaminated each other. C , Anterograde labeling in the IO following transsynaptic tracing; the labeling is predominantly ipsilateral to the MCN ( Ci ) and LCN ( Cii ) that were injected. Note that some transsynaptically labeled cells can be observed in the IO contralateral to MCN injection spot ( Ci ). D , Images of a fiber in the caudal MAO with varicosities (yellow arrows) positive for mCherry ( Di ), GAD67 ( Dii ), or both ( Diii ). E , Number of GAD67+ and GAD67− varicosities in cdMAO for MCN and LCN injections. Error bars represent standard deviation (SD), and dots represent number of varicosities in individual mice. F , Summary panel demonstrating the laterality of the observed inhibitory disynaptic projection from the MCN to the cdMAO and the complementary direct excitatory projection from the MCN.

Article Snippet: For the transsynaptic experiments in wild types, we used adeno-associated virus AAV1-CMV-HI-eGFP-Cre-WPRESV40 (titer 8 × 10 12 vg/ml, 105545-AAV1; Addgene) that expresses Cre and GFP in the nucleus, while applying AAV9-hsyn-DIO-mCherry (>1 × 10 13 vg/ml, No 50459-AAV9; Addgene) in the SC as a Cre-dependent tracer.

Techniques: Expressing, Injection, Labeling, Standard Deviation