Transcriptional profiling by array to study the effect of adenoviruses encoding the FLAG epitope or full-length antisense sequence to Atf3 on transcriptional regulation in rat cardiac myocytes

ABSTRACT: Cardiomyocytes, the terminally-differentiated contractile cells of the heart, undergo hypertrophic growth in response to endothelin-1. This is associated with changes in mRNA expression of immediate early genes (IEGs). Atf3 is a member of the ATF/CREB family transcription factors which bind as dimers to the cAMP response element (CRE). Atf3 mRNA is particularly associated with cell stress responses and is implicated in negative feedback regulation of gene expression. We confirmed that endothelin-1 promotes expression of Atf3 mRNA and protein in cardiomyocytes and that it is an IEG. Atf3 expression was transient and maximal at 0.5-1 h. We used adenoviruses for expression of antisense Atf3 RNA in cardiomyocytes to inhibit upregulation of Atf3 protein in response to endothelin-1. The effects on the cardiomyocyte transcriptome in control cells and cells exposed to endothelin-1 were examined using Affymetrix rat exon 1.0 ST arrays with data analysis using GeneSpring GX11.5. We identified transcripts that were upregulated by endothelin-1 for which antisense Atf3 significantly (FDR<0.05) enhanced or inhibited expression, respectively. The basal levels of other transcripts that are upregulated by endothelin-1 were also increased by antisense Atf3, though antisense Atf3 had no further effect on the response to endothelin-1. The data were validated by quantitative PCR of selected IEG mRNAs examining the effects at different times.

ORGANISM(S): Rattus norvegicus

SUBMITTER: Angela Clerk 

PROVIDER: E-MEXP-3392 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress