Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of Drosophila wild type Canton S vs. otd-uvi eye specific mutant alles of Otd revals regulation of gene networks in photoperception-related functions by the CRX/OTX homologue

ABSTRACT: Comparative global gene expression analysis was caried out using pupal head tissue of otd-uvi an eye specific mutant allele of Otd and wild type control Canton S., in order to delineate the function of homeobox transcription factor Otd in Drosophila photoreceptor development and function. Experiment Overall Design: The Drosophila strain Canton S was used as wild type for comparative gene expression analysis with the mutant strain otd-uvi. For each biological replicate, samples were derived from independent RNA extractions consisting of 15-20 pupal heads. Pupae were dissected at 80 hr APF and immediately transferred to 600ul of trizol (GIBCO/BRL). RNA extraction was performed according to manufacturers instructions. Qualitative and quantitative analysis of the total RNA was carried out by capillary electrophoresis on a RNA6000 Nano Assay chip, with Agilent 2100 Bioanalyzer (Agilent Technologies USA) Biotin labeled probes were generated from 10ug total RNA according to Affymetrix protocol.

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Francesca Pignoni

PROVIDER: E-GEOD-5321 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Analysis of the Otd-dependent transcriptome supports the evolutionary conservation of CRX/OTX/OTD functions in flies and vertebrates.

Ranade Swati S SS Yang-Zhou Donghui D Kong Sek Won SW McDonald Elizabeth C EC Cook Tiffany A TA Pignoni Francesca F

Developmental biology 20080131 2

Homeobox transcription factors of the vertebrate CRX/OTX family play critical roles in photoreceptor neurons, the rostral brain and circadian processes. In mouse, the three related proteins, CRX, OTX1, and OTX2, fulfill these functions. In Drosophila, the single founding member of this gene family, called orthodenticle (otd), is required during embryonic brain and photoreceptor neuron development. We have used global gene expression analysis in late pupal heads to better characterize the post-em ...[more]

PMID: 18241855

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Project description:Background Many microarray experiments search for genes with differential expression between a common “reference” group and multiple test groups, like in the case of time-course designs or of various treatments versus a control condition. In such cases, currently employed statistical approaches based on t-test or close derivatives have limited efficacy, mostly because estimation of noise is done on only two groups at time. Alternative approaches based on ANOVA correctly capture noise from all the groups, but then do not confront single test groups with the reference. We therefore conceived a statistical test for pairwise comparisons between the reference group and each test group that uses within-group variance calculated from all the groups. Results We implemented an R-Bioconductor package named Mulcom, with a statistical test derived from the Dunnett’s test, designed to compare multiple experimental groups against a common reference. In addition to the basic Dunnett’s t value, the package includes an optional minimal fold-change threshold, m. Thanks to automated, permutation-based estimation of False Discovery Rate (FDR), the package also permits fast optimization of the test, to obtain the maximum number of significant genes at a given FDR value. When applied on a time-course experiment profiled in parallel on two microarray platforms, and compared with currently used tests, Mulcom displayed higher concordance of significant genes in the two array platforms, and higher enrichment in functional annotation to categories related to the biology of the experiment. Conclusions The Mulcom package provides a fast and powerful tool for the identification of differentially expressed genes when several experimental conditions are compared with a common reference. We found that Mulcom leads to lists of differentially expressed genes that are particularly consistent across microarray platforms and enriched in significant classes of genes. In our opinion, the main reasons for these good performances are three: (i) within-group variability is estimated from all experimental groups even if only two of them are compared each time; (ii) the optional fold-change threshold m avoids false positives due to aberrantly low within-group variability; (iii) automated test optimization allows maximizing sensitivity without compromising specificity. Ten MDA-MB-435 samples, biological duplicates of each condition (untreated, integrin Beta4 treatment, hepatocyte growth factor treatment for 1 hr, 6 hrs, or 24 hrs).

Dataset Information

Transcription profiling of Drosophila wild type Canton S vs. otd-uvi eye specific mutant alles of Otd revals regulation of gene networks in photoperception-related functions by the CRX/OTX homologue

Publications

Analysis of the Otd-dependent transcriptome supports the evolutionary conservation of CRX/OTX/OTD functions in flies and vertebrates.

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