Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptional profiling of the ?mak-1 strain

ABSTRACT: The MAK-1 pathway is the least characterized MAP kinase pathway in Neurospora. Microarray analyses were carried out to identify genes with altered expression in ?mak-1 as compared to wild type strains. Mycelial mats of wild type and ?mak-1 strains were grown in constant light (LL) at 25C for 2 and 6 days, respectively, in 1X Vogel’s, 2% glucose, 0.5% Arginine HCl, pH 6.0. A cork borer was used to cut plugs to inoculate 75mL of the same liquid media and cultures were grown in LL at 25C for 24h, transferred to constant darkness (DD) 25C for 24 h before being harvested and total RNA extracted. One-condition experiment of the ?mak-1 mutant strain compared to the wild type strain. Cy3 and Cy5 dye swaps were performed.

ORGANISM(S): Neurospora crassa

SUBMITTER: Lindsay Bennett

PROVIDER: E-GEOD-41778 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Background: The incidence of alcohol and tobacco co-abuse is as high as 80%. The molecular mechanism underlying this comorbidity is virtually unknown but interactions between these drugs have important implications for the development of, and recovery from drug dependence. Methods: We investigated the effects of chronic tobacco and alcohol abuse and the interaction of the two behaviours on global gene expression in the human nucleus accumbens using cDNA microarrays and 20 alcoholic and control cases, with and without smoking comorbidity. Changes in gene expression were established by two-way ANOVA. Unsupervised hierarchical clustering was utilized to probe the strength of the data sets. Results: Subjecting the data sets derived from microarray gene expression screening to unsupervised hierarchical clustering tied the cases into distinct groups. When considering all alcohol-responsive genes, alcoholics were separated from non alcoholics with the exception of one control case. All smokers were distinguished from non smokers based on similarity in expression of smoking-sensitive genes. In the nucleus accumbens, alcohol-responsive genes were associated with transcription, lipid metabolism and signalling. Smoking-sensitive genes were predominantly assigned to functional groups concerned with RNA processing and the endoplasmic reticulum. Both drugs influenced the expression of genes involved in matrix remodelling, proliferation and cell morphogenesis. Additionally, a gene set encoding proteins involved in the canonical pathway ‘regulation of the actin cytoskeleton’ was induced in response to alcohol and tobacco co-abuse and included. Conclusions: The region-specific modulation of alcohol-sensitive gene expression by smoking may have important consequences for alcohol-induced aberrations within the mesolimbic dopaminergic system. The data set contained individual RNA samples extracted from the nucleus accumbens from 5 non smoking non drinking control case, 5 non smoking alcoholics, 5 non drinking smokers, and 5 smoking alcoholics. An aliquot if each sample was combined to generate a reference pool. Each sample was amplified individually and labelled with Cy3 in the process, the reference samples was amplified and labelled with Cy5. Each sample was hybridised competitively against the reference sample to 1 array.

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Transcriptional profiling of the ?mak-1 strain

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