VAMP2 sequestration by α-syn aggregates in cultured primary neurons results in synaptic dysfunction as well as cytotoxicity.

<p>(A) Immunocytochemistry in rat cortical primary neurons shows a direct binding of VAMP2 to α-syn aggregates. (B) Western blot analysis of cultured neurons shows selectively reduced VAMP2 and SNAP25 levels in α-syn aggregates-treated cells for 3 days consecutively. To analyze bound VAMP2, the blot used for α-syn aggregates detection was re-probed, marked in the red box. (C) Cortical neuronal cells were treated with α-syn aggregates for 3 days consecutively and glutamate release was determined. ****, p < 0.0001, by two-tailed, unpaired <i>t</i>-test (n = 10). (D) Cell viability was determined 14 days after addition of α-syn aggregates in cortical neurons. ***, p < 0.001, by two-tailed, unpaired <i>t</i>-test (n = 3). Values indicate mean ± SEM.