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Kc167_Inter-slide_Variability_Test

ABSTRACT: A large batch of Drosophila Kc167 total RNA was prepared and was used for 3 homotypic hybridizations in order to evaluate the inter-slide variability using the CDMC_Drosophila_7k2 platform. See Neal et al. 2003 for results and discussion. Keywords = Kc167 Keywords = Drosophila Keywords = Platform Variability

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Meredith Gibson

PROVIDER: E-GEOD-418 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Construction of a cDNA-based microarray for Drosophila melanogaster: a comparison of gene transcription profiles from SL2 and Kc167 cells.

Neal Scott J SJ Gibson Meredith L ML So Anthony K-C AK Westwood J Timothy JT

Genome 20031001 5

We have constructed a DNA microarray that represents approximately 6900 of the estimated 13,598 genes in the Drosophila melanogaster genome. The microarray contains 5756 target cDNAs from the Berkeley Drosophila Genome Project, 1078 cDNAs from the National Institutes of Health Drosophila testis cDNA library, and 546 gene fragments that were amplified from genomic DNA. The methods for DNA amplification and microarray manufacture are presented. Academic researchers can obtain the microarray from t ...[more]

PMID: 14608405

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SL2_vs_Kc167_Dif_Expr

Project description:The expression profiles of Drosophila Kc167 and SL2 cells were compared on the CDMC_Drosophila_7k2 array platform. Three independent RNA samples from each cell line were isolated and compared in a pairwise fashion. Dye-flip experiments were also performed. See Neal et al. 2003 for results and discussion. Keywords = Kc167 Keywords = SL2 Keywords = Drosophila

2010-06-05 | E-GEOD-419 | biostudies-arrayexpress

Effects of mutation in a soluble guanylyl cyclase gene

Project description:Genomic and behavioral investigations were performed to determine the effects of a mutation in a Drosophila soluble guanylyl cyclase gene. A mutant DGCalpha1[3] third chromosome was crossed into a natural rover (for[R]) or natural sitter (for[s]) genetic background. (See Osborne et al. 1997; PMID: 9242616.) First instar larvae were collected and grown on 60mm Petri plates containing 10 mL of food until mid-third instar. (Approximate density was 3 animals per mL food). Larvae were collected and washed quickly with distilled water and were flash frozen in liquid nitrogen. Co-reared larvae were tested for behavioural effects. Four independent collections were made for each of the two conditions (Rover_DGCalpha1[3] or sitter_DGCalpha1[3]). Keywords = Drosophila Keywords = foraging Keywords = behavior Keywords = cGMP Keywords = guanylyl cyclase Keywords = genetic background

2010-06-10 | E-GEOD-2136 | biostudies-arrayexpress

Thermoprotection in the Drosophila hsf4 mutant

Project description:In Drosophila larvae, acquired synaptic thermotolerance following heat shock has previously been shown to correlate with the induction of heat shock proteins (Hsps) including HSP70. We tested the hypothesis that synaptic thermotolerance would be significantly diminished in a temperature-sensitive strain (hsf4) which has been reported not to be able to produce inducible Hsps in response to heat shock. Contrary to our hypothesis, considerable thermoprotection was still observed at hsf4 larval synapses following heat shock. To investigate the cause of this thermoprotection, we conducted DNA microarray experiments to identify heat-induced transcript changes in these organisms. Transcripts of the hsp83, dnaJ-1(hsp40) and gstE1 genes were significantly up-regulated in hsf4 larvae after heat shock. In addition, increases in the levels of Hsp83 and DnaJ-1 proteins but not in the inducible form of Hsp70 were detected by Western blotting. The mode of heat shock administration differentially affected the relative transcript and translational changes for these chaperones. These results indicate that the compensatory up-regulation of constitutively expressed Hsps, in the absence of the synthesis of inducible Hsps including HSP70, could still provide substantial thermoprotection to both synapses and the whole organism. Two strains were used in this study; the mutant cn bw hsf4 strain, and the dp cn bw cl control strain from which the mutant was derived. For each strain, co-reared larvae were separated into a control group and a treatment group at random. RNA from the heat-shocked larvae and from the respective controls were directly compared on cDNA microarrays (GPL311). Two independent biological samples from each genotype were collected. In addition, an inter-strain comparison was made using the RNA from the untreated controls from each genotype. This was also performed in duplicate.

2005-11-30 | E-GEOD-2998 | biostudies-arrayexpress

Cold hardening gene expression

Project description:Cold hardening treatment, a brief exposure to low temperatures (e.g. 0°C for 2 h), can protect certain insects against subsequent exposure to temperatures sufficiently low to cause damage or lethality. Microarray analysis to examine the changes in transcript abundance associated with cold hardening has been undertaken in Drosophila melanogaster in order to gain insight into this phenomenon. Transcripts associated with 36 genes were identified, a subset of which appeared to be also differentially expressed after heat shock treatment. Quantitative RT-PCR was used to independently determine transcript abundance of a subset of these sequences. Taken together, these assays suggest that stress proteins, including Hsp23, Hsp26, Hsp83 and Frost as well as membrane-associated proteins may contribute to the cold hardening response. Co-reared flies were separated into a control group and a treatment group at random. RNA was isolated from the cold-shocked flies and the respective controls and was used for direct comparisons on cDNA microarrays. Treatments were not varied as they had been optimized in previous studies. A pilot experiment was performed using the Drosophila 7k2 array (GPL311) and subsequently three pairs of independent biological samples were evaluated with the Drosophila 12k1 array (GPL1467).

2005-11-30 | E-GEOD-2989 | biostudies-arrayexpress

Effects of methotrexate on S3 Drosophila cells and Drosophila ovaries

Project description:Comparison of transcript abundance between control (untreated) and methotrexate treated S3 Drosophila cells and ovaries disected from female flies. S3 cells were exposed to 0 or 5.2 X 10^-8 M MTX for 4 days then harvested. Flies were exposed to 0 or 5 ppm MTX for 5 days then ovaries were dissected. RNA was extracted from S3 cells and ovaries. 4 S3 microarrays with one dye-swap and 2 ovary microarrays swapping dyes were hybridized to spotted cDNA microarrays.

2010-06-30 | E-GEOD-3495 | biostudies-arrayexpress

Drosophila gene expression in response to the botanical insecticide Piper nigrum

Project description:This experiment was used to determine the effect of a botanical insecticide upon gene expression profiles in Drosophila melanogaster. Adult female Drosophila (oregon-R strain) were treated with an ethylacetate extract of Piper nigrum (Piperaceae) seeds formulated in 99% ethanol. Treatment was topical, using a Potter's tower to administer a total of 2 mL of a 0.9mg/mL concentration. Control treatment was identical except flies were treated with 99% ethanol as a solvent control. Gene expression was studied four hours post-treatment. The design is a direct comparison between total RNA of treated and untreated insects. Sample size is two and two reverse label hybridizations were done. Each sample is a pool of 240 insects which received the same treatment simultaneously.

2006-01-24 | E-GEOD-3032 | biostudies-arrayexpress

Evalutation of the effect of ethanol upon gene expression in adult Drosophila melanogaster

Project description:This experiment was intended to determine if topical exposure to ethanol led to significant changes in gene expression in adult Drosophila melanogaster. It was determined that topical exposure to ethanol did not lead to a strong differential expression of genes in Drosophila after 4 hours. The effect of topical treatment with 99% ethanol was compared to the effect of a water treatment which was used as a control and the differential expression between these two treatments was assessed four hours after treatment. Two biological replicates of each treatment were compared for a total of two microarray hybridizations

2010-06-11 | E-GEOD-3033 | biostudies-arrayexpress

Gene expression profiling in hepatic tissue of newly weaned pigs fed pharmacological zinc supplemented diets

Project description:Zinc (Zn) is an essential trace element for all life forms. Zn supplementation has been used to treat diarrheal disease in children, and in the U.S. swine industry at pharmacological levels to promote growth and fecal consistency, but underlying mechanisms explaining these beneficial effects remain unknown. Thus, we hypothesized that the benefits of pharmacological Zn supplementation were a result of changes in gene expression. For this study, liver RNA from newly weaned pigs fed dietary Zn as Zn oxide for 14 days at either adequate (150 Zn/kg) or pharmacological (2000 mg Zn/kg) levels was evaluated using a 70-mer oligonucleotide microarray. Interrogation of this microarray revealed 658 annotated transcripts (FDR ≤ 0.05) affected by pharmacological Zn supplementation. Relative real-time RT-PCR was used to confirm differential expression of two genes. Results suggest that feeding pharmacological Zn (2000 mg Zn/kg) affects genes involved in reducing oxidative stress and in amino acid metabolism, which are essential for cell detoxification and proper cell function. Oligonucleotide microarrays used for this study consisted of 13,297 70-mer oligos (Pig Array-Ready Oligo Set v. 1.0 and Pig Oligo Extension Set v. 1.0, Qiagen, Inc., Valencia, CA) each spotted once on a single slide. Slides were printed at the Michigan State University Research Technology Support Facility. Oligonucleotides spotted in multiple locations for use as potential controls included 76 Arabidopsis thaliana gene spots, 17 beta tubulin spots, 17 glyceraldehyde-3-phosphate dehydrogenase spots, 85 heat shock protein gene spots, 69 ribosomal protein gene spots, 112 randomly generated negative control spots and 470 blanks. The microarray was screened with the Zn150 and Zn2000 samples, and four microarray slides were screened. Zn150 samples were randomly paired with Zn2000 samples. Two samples from each treatment were labeled with Cy3 and the other two were labeled with Cy5.

2008-10-02 | E-GEOD-11972 | biostudies-arrayexpress

267B1, PC3M, LNCaP Methylation Profile vs. mRNA Expression Level

Project description:Two prostate immortalized cell lines, 267B1, RWPE-1, Ml-csv40, and five prostate cancer cell lines, PC3, PC3M, PC3M-Pro4, PC3M-LN4 and LNCaP are included in this experiment. Their methylation profile are detected using human promoter array (v11). mRNA expression level are profiled with Affymetrix U133A Arrays for 267B1 and PC3M. Keywords: other

2010-06-09 | E-GEOD-951 | biostudies-arrayexpress

Serum Starvation/Refeeding RNA vs. RAP

Project description:Low abundance mRNAs are more difficult to examine using microarrays than high abundance mRNAs due to the effect of concentration on hybridization kinetics, signal-to-noise ratios, and interference between signals from mRNA isoforms. Individual sequences in LCRs are more highly represented than in the mRNA population from which they are derived, leading to favorable hybridization kinetics. LCR targets permit the measurement of abundance changes that are difficult to measure using oligo dT priming for target synthesis. An oligo dT-primed target and three LCRs detect twice as many differentially regulated genes as could be detected by the oligo dT-primed target alone, as serum-starved fibroblasts responded to the reintroduction of serum. Thus, this target preparation strategy considerably increases the sensitivity of spotted cDNA microarrays.

2010-06-10 | E-GEOD-2655 | biostudies-arrayexpress

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