Tetanus Neurotoxin (TeNT) enters the peripheral endings of motor axons in muscles and transports to motoneuron cell bodies where transynaptically translocates to presynaptic inputs blocking inhibitory synapses. This disinhibits motoneurons causing dysregulated firing and tetanic contractions. Preferential actions on inhibitory synapses are usually explained by enrichment in the Vesicle-Associated Membrane Protein 2 isoform (VAMP2). VAMP1 and VAMP2 form part of the SNARE complex and are differentially distributed in different synapses. Nevertheless, the exact VAMP content inside synapses on motoneuron cell bodies has never been examined (except for cholinergic C-terminals enriched in VAMP2). We used immunocytochemistry to examine VAMP1 and VAMP2 in postsynaptic inhibitory synapses (marked by vesicular GABA/Glycine transporters, VGAT) and in excitatory synapses marked by vesicular glutamate transporters: VGLUT1, proprioceptive synapses; VGLUT2, synaptic inputs from spinal interneurons and descending projections. Analyses were done on Fast Blue retrogradely labeled gastrocnemius motoneurons. We also examined VAMP content in P-boutons (GAD65+) presynaptic to VGLUT1 synapses. Inhibitory synapses contained more VAMP1 than VAMP2, while P-boutons only express VAMP2. VGLUT1 boutons contain only VAMP1 and VGLUT2 boutons express both VAMPs. To examine VAMP changes after TeNT we injected different doses in muscle to elicit a mild tetanus. We found that both VAMP1 and 2 decreased inside inhibitory synapses, with a larger effect on VAMP2. However, VAMP1 was unaffected in VGLUT1 and VGLUT2 synapses, and VAMP2 barely diminished in VGLUT2 boutons. Thus, TeNT actions are not explained by VAMP isoform content, but more likely by differential entry in inhibitory vs excitatory synapses affecting both VAMPs.