Journal: PLoS ONE
Article Title: Torsin A Localization in the Mouse Cerebellar Synaptic Circuitry
doi: 10.1371/journal.pone.0068063
Figure Lengend Snippet: Western blot of TA, V1, V2, VGAT proteins in cerebellar synaptosomal fractions (SYN) from P14 mice. Synaptosomal preparations are enriched of post synaptic density protein (PSD95) and Synapsin-I (Syn-I) in comparison to equal amount (ACT) of total cerebellar protein extract (LYS). TA is clearly present in synaptosomal enriched preparations. B ) Cerebellar sagittal section immunostained for Cb (green), V1 (cyan) and TA (red). In the ML, anti-V1 specifically labels PF-terminals contacting PC-spines. C – E ) The high magnification of the inset in B is reproduced in different merge images and by the relative split channels. C ) Merge image of Cb and V1 staining plus the colocalization mask Cb/V1 (mCb/V1 in white); it represents the negative CTR (CTR-). D ) Merge image of TA and Cb staining plus the colocalization mask TA/Cb (mTA/Cb in white), highlighting TA expression in PC-spines. E ) Merge image of TA and V1 staining plus the colocalization mask TA/V1 (m TA/V1 in white), indicating the presence of TA also in this synaptic terminal. F ) Quantitative colocalization analysis shows the mean overlap coefficients of TA/Cb (gray column) and TA/V1 (black column) significantly different from the negative CTR Cb/V1 (white column). The small insets (white boxes in C–D-E) are high magnifications of a representative PC-spine (green) contacted by a PF synaptic terminal (cyan) with the appropriate colocalization mask (white). * p < 0.05. Data are represented as mean ± SEM. Scale bars in B, 10 µm; in C–D-E, 5 µm; in F, 1 µm.
Article Snippet: 131011), rabbit anti-Synapsin-I 1:1000 for 1 ON (Novus Biologicals NB300-104), mouse anti-post synaptic density 95 protein (PSD95) 1:20000 1h (Millipore, cod.
Techniques: Western Blot, Staining, Expressing