Journal: PLoS ONE

Article Title: Neuroligin-1 Loss Is Associated with Reduced Tenacity of Excitatory Synapses

doi: 10.1371/journal.pone.0042314

Figure Lengend Snippet: A ) Distribution of range over mean values for SEpH:GluA2 puncta in the presence of CNQX (10 µM), AP-5 (50 µM) and TTX (1 µM) (Nlgn-1 KO: 10 neurons, 481 synapses; WT: 10 neurons, 514 synapses). B ) SI decay rates in the presence of the aforementioned pharmacological agents.

Article Snippet: Reagents were procured from the following sources: CNQX (6-cyano-7 nitroquinoxaline-2,3-dione) from Tocris Bioscience; AP-5 (2-amino-5-phosphonopentanoic acid) from Sigma-Aldrich; TTX (tetrodotoxin) from Alomone Labs. A mixture of CNQX, AP-5, and TTX were diluted in 100 µl medium drawn from the cloning cylinder in which the neurons were grown.

Techniques:

Journal: PLoS ONE

Article Title: Enrichment of Conserved Synaptic Activity-Responsive Element in Neuronal Genes Predicts a Coordinated Response of MEF2, CREB and SRF

doi: 10.1371/journal.pone.0053848

Figure Lengend Snippet: Upregulation of SARE identified candidates in primary cortical neurons. Primary neurons from E18 cortex were cultivated for 16 days before triggering neuronal activation with 4AP/bicuculline. RNA was obtained and transcript expression of SARE containing genes analyzed by quantitative real time RT-PCR (Q-PCR). Expression of each gene is normalized to the expression in control cells. Arc up-regulation demonstrated efficient neuronal activation. * indicates p<0,5 **<0,1, ***<0,05 (t-Student test). B). Novel SARE sequences activate transcription in response to neuronal activity. Mouse genomic sequence containing SARE regulatory regions (see below) corresponding to those identified for the Cux1 and Cux2 genes were cloned into the pGL4.23 luciferase vector (Promega). Neurons obtained from E18,5 cortex were co-transfected with the indicated firefly luciferase reporter construct and internal control Renilla luciferase plasmid at a ratio of 4∶1. Neuronal activity was trigered with 4AP/bicuculline before measuring transcription of the reporters. Relative expression of each reporter constructs was determined by normalizing the activity of each reporter to its activity on TTX treated neurons. Data represent mean and standar deviation of results obtained in three different experiments. * indicates p<0,01, **<0,001 (t-Student test).

Article Snippet: 12 h before inducing neuronal activity cells were incubated with 2 mM tetrodotoxin (TTX) (Alomone-labs, Jerusalem, Israel).

Techniques: Activation Assay, Expressing, Quantitative RT-PCR, Activity Assay, Sequencing, Clone Assay, Luciferase, Plasmid Preparation, Transfection, Construct