01 were considered for the analysis. Right-tailed Fisher’s exact test was used to calculate a p value determining the probability that each biological function

and/or disease assigned to that data set is due to chance alone. Two milliliters of shaved synaptosomes collected from sucrose gradients were diluted in 5 ml PBS and centrifuged for 30 min at 5,500 × gmax, 4°C, in a swing out rotor. The synaptosomal pellet was then resuspended in 2.4 ml PBS. One hundred microliters of this suspension were carefully aliquoted onto each poly-L-lysine precoated coverslip placed in a 12-well plate and incubated for 45 min at room temperature. Afterward, 1 ml PBS was selleck compound added to each well and synaptosomes pelleted on the coverslip by centrifugation for 30 min at 5,580 × gmax (5,500 rpm) at 4°C in a HIGHplate rotor Epacadostat supplier 75006444 (Sorvall Heraeus). Synaptosomes were then fixed with 4% paraformaldehyde (PFA), permeabilized with 0.1% Triton X-100 and stained with primary and secondary antibodies. Synaptosomal images were acquired using an AOBS SP2 confocal microscope (Leica Microsystems) with a 63× oil-immersion objective, standard filter sets (Leica Microsystems), and Leica LCS software. Line scan analyses

were performed using the LAS AF Lite software (Leica). The extent of colocalization between different protein pairs in glutamaterigic versus GABAergic synaptosomes were determined using a custom written Matlab algorithm (The Mathworks Inc.) kindly provided by Prof. Silvio Rizzoli. Colocalization with either enough VGLUT1 or VGAT was considered when the center of intensity in

the two channels was within a distance of 200 nm. At least 500 synaptosomes were analyzed for each given protein pair in three independent biological replicates. We thank Prof. Silvio Rizzoli for providing the MatLab program and Maria Druminski for excellent technical assistance. The research leading to these results has received funding from the European Union Seventh Framework Programme under grant agreement no. HEALTH-F2-2009-241498 (“EUROSPIN”) to R.J. and of the Deutsche Forschungsgemeinschaft (SFB 889, TP4) to R.J. and H.U. “
“The formation of long-lasting memory requires de novo synthesis of mRNA and proteins and is blocked by inhibitors of transcription or translation, whereas short-term memory relies on the modification of pre-existing proteins and is not affected by such inhibitors. Similarly, long-term potentiation (LTP), a cellular model for learning and memory, is dependent on transcription and translation for its late phase (late LTP; L-LTP), which lasts for many hours, while its early phase (early LTP; E-LTP) lasts 1–2 hr and is translation independent (Kandel, 2001; Silva, 2003). Neuronal mRNA translation is tightly regulated by synaptic activity (Banerjee et al., 2009; Kelleher et al., 2004a; Richter and Klann, 2009; Sutton and Schuman, 2006).