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E-GEOD-22467 - Transcription profiling by array of human SK-N-AS neuroblastoma cells treated with constructs expressing truncated or full-length NRSF
Released on 30 July 2010, last updated on 1 May 2014
Neuron-restrictive silencer factor (NRSF) and its isoforms are differentially regulated in rodent models of self-sustaining status epilepticus (SSSE). NRSF isoforms regulate genes associated with SSSE, including the proconvulsant tachykinins, brain-derived neurotrophic factor and multiple ion channels. NRSF isoforms may direct distinct gene expression patterns during SSSE and the ratio of each isoform may be a causative factor in traumatic damage to the CNS. Here we analysed global gene expression changes by microarray in human SK-N-AS neuroblastoma cells following the over expression of NRSF and a truncated isoform, HZ4. We used bioinformatics software to analyse the microarray dataset and correlated these data with epilepsy candidate gene pathways. Findings were validated by RT-PCR. We demonstrated that NRSF and HZ4 direct overlapping as well as distinct gene expression patterns and that they differentially modulated gene expression patterns associated with epilepsy. Finally we revealed that NRSF gene expression may be modulated by the anticonvulsant, phenytoin. This study provides fundamental information on networks of genes that may be altered during SSSE, following altered NRSF expression, which may be important in future therapeutic research and clinical analysis of genetic variation predisposing to epilepsy. Human SK-N-AS cells were treated with expression constructs over-expressing either the full length human transcription factor, NRSF, (via the RE-EX1 construct) or a truncated variant (via the HZ4 construct). Cells were treated for either 0hrs (base line controls), 24hrs or 48hrs, before being immediately processed for RNA extraction. An affymetrix microarray was employed to investigate gene expression patterns, comparing each time point (24hrs or 48hrs) againsts its baseline ()hrs) control. This experiment was performed in triplicate samples per batch, and over three batches.
transcription profiling by array
Stuart Gillies <email@example.com>, B Von Mentzer, G Jacobson, J P Quinn, K Haddley, S G Gillies, S Vasiliou, V J Bubb