Fig 1: (A) Daily expression of Synapsin 2 (SYN2) and Vesicle-fusing ATPase (NSF) in the cerebellum, assessed by two-dimensional difference in gel electrophoresis (2D-DIGE; left panels; n = 4–6 per ZT) and western blots (WB; right panels; n = 5 per ZT). Fitted curves represent significant cosinor regressions. A.U., arbitrary unit. Data are presented as mean ± SEM. (B) Representative western blots for SYN2 and NSF. ZT, Zeitgeber time.
Fig 2: Syn II expression rescues the epileptic phenotype observed in Syn2 KO animals. (A) Seizure activity as a function of age for WT (black), Syn 2 KO (red), and AAV1.hSyn.Syn2-Turbo-GFP-injected Syn2 KO rescue (blue). The rescue is almost complete at P15 and it weakens as the postnatal age increases. (B) Seizure activity at P15 including mock-injected GFP controls (AAV1.hSyn.Turbo-GFP-injected Syn2 KO). n = 9 for each group. * p < 0.05, ** p < 0.01, and *** p < 0.001 per ANCOVA followed by Tukey’s test (injected versus Syn2 KO).
Fig 3: Phosho-incompetent Syn2S10A mutant partially rescues the overexcitable phenotype in Syn2 KO animals. (A) A diagram showing the conserved domains (A–C) of Syn II and the PKA/CaMKI phosphorylation site P1 in the A domain. The diagram below illustrates Syn II (red) function in tethering and clustering of synaptic vesicles and directing them to Ca2+ channels (yellow). (B) ICV of the native Syn2 and mutant Syn2S10A genes produce a similar reduction in spontaneous synaptic activity (n = 16). (C) Expression of mutant Syn2S10A does not reduce the 4-AP produced epileptiform activity to the extent observed for the expression of native Syn2 (p < 0.05, n = 9). (D) Expression of mutant Syn2S10A does not reduce seizure activity to the extent observed for the expression of native Syn2 (* p < 0.05, n = 9). Dotted red lines corrrespond to Syn2 KO animals, and dotted black lines correspond to WT animals (replotted from Figure 3, Figure 4 and Figure 5).
Fig 4: Expression of SynII-TurboGFP in the hippocampus of AAV-injected mice. (A) SynII-TurboGFP expression in the hippocampus and cortex after neonatal ICV injection of AAV1.hSyn.Syn2-Turbo-GFP. Scale 500 µm. (B) High-magnification image of a small section of the Str. Radiatum showing the punctate immunoreactivity expected for SynII-TurboGFP. Scale 50 µm. (C) The distributions of the anti-Syn II fluorescence for the pixels colocalized (pink) and not colocalized (navy) with the anti-GFP signal (green channel). Colocalization was assessed using ImageJ plugin Coloc 2. Note that the non-colocalized signal matches the background peak observed for Syn2 KO slices (shown in Figure 1C), while the colocalized peak matches the anti-Syn II signal observed in WT slices (p < 0.001 per K-S test).
Fig 5: Neonatal Syn2 expression rescues the epileptiform activity observed upon 4-AP application (30 µM) in Syn2 KO hippocampal slices. (A) Representative recordings from WT, Syn2 KO, and AAV1.hSyn.Syn2-Turbo-GFP-injected Syn2 KO animals with 30 µm 4-AP applied to the bath. Arrow denotes an I-IC event [31]. (B) I-IC spikes are induced in Syn2 KO slices upon 4-AP application, and this hyperexcitability is rescued in AAV1.hSyn.Syn2-Turbo-GFP-injected animals. * p < 0.05 per ANOVA followed by Tukey’s test, n = 9.
Supplier Page from Abcam for Anti-SYN2 antibody