Expression differences between inversions and their progenitors in Drosophila melanogaster
Ontology highlight
ABSTRACT:
ORGANISM(S): Drosophila melanogaster
SUBMITTER: Steven Russell
PROVIDER: E-GEOD-16616 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Having developed a well characterized acquired model of behavioral and gene expression plasticity, we found an opportunity here to investigate if drug exposure to adult males could cause transgenerational effect. To probe this, we carried out a systematic study at both behavioral and microarray gene expression levels. In the latter, we asked the question that do F0 testis, F1 males’ head, F1 females’ head, F1 testis, F2 males’ head and F2 females’ head show gene expression changes if F0 male parents had a history of PTZ exposure? A total of 28 microarray slides were used in this study. Unless mentioned otherwise, standard method of fly handling and manipulation was used. Freshly generated isogenic D. melanogaster Oregon-R wild type flies were grown in standard fly medium consisting of agar-agar, maize powder, brown sugar, dried yeast, and nipagin. The cultures were grown at 24 + 1oC, 60% RH, and 12 hrs light (9 AM to 9 PM) and 12 hours dark cycle. Three to four days old unmated adult males were grown in food containing 8 mg/ml PTZ for seven days. Flies were then shifted to vials containing NF. Seven days after shifting, F0 males were mated to same age group females maintained on NF throughout, without any drug exposure. Males and females were housed in groups in vials containing NF. Eggs collected on NF were used to generate F1 individuals as unmated males and virgin females. F1 males were mated to same age group females which were independently collected and maintained on NF. These females had no history of drug exposure in prior generations. Males and females were housed in groups in vials containing NF. Eggs collected on NF were used to generate F2 individuals as unmated males and virgin females. F0 testis, F1 males’ head, F1 females’ head, F1 testis, F2 males’ head and F2 females’ head were used for RNA isolation. Total RNA was isolated from pools of heads or testis, using TRI REAGENT (Sigma) according to the manufacturer’s protocol. Double stranded cDNA was synthesized from extracted RNA using Microarray cDNA Synthesis Kit (Roche). The cDNA was purified using Micorarray Target Purification Kit (Roche), according to the manufacturer’s protocol. Each of the four sets of control (no PTZ exposure of F0 males) and treated cDNA samples, belonging to the four biological replicates, was used for labeling with either Cy3 or Cy5 dyes (Amersham Biosciences) using Microarray RNA Target Synthesis Kit T7 (Roche). The labeled products were purified by Microarray Target Purification Kit (Roche). The Cy3 and Cy5 labeled two cRNA samples of each biological replicate were pooled together, precipitated, washed, air-dried, and dissolved in 18MΩ RNAase free water (Sigma). 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