Project description:Deep sequencing of total RNA extracted from the genital discs of males for each of the following strains : Drosophila sechellia, Drosophila mauritiana, hybrid introgression line 3Q1(A) and hybrid introgression line Q1(A)
Project description:Deep sequencing of total RNA extracted from the genital discs of males for each of the following strains : Drosophila sechellia, Drosophila mauritiana, hybrid introgression line 3Q1(A) and hybrid introgression line Q1(A) Analysis of poly(A)+ RNA for three independent biological replicates of sequencing libraries for each of the following strains: D. sechellia w, D. mauritiana P-insertion Q1, hybrid introgression line 3Q1(A), and hybrid introgression line Q1(A). Male genital discs were obtained as described above, and total RNA was extracted using RNAqueousM-CM-^BM-BM-.-Micro Kit (Ambion). Poly(A)+ transcripts were isolated subsequently using MicroPoly(A)PuristM-CM-"M-BM-^DM-BM-" Kit (Ambion). To facilitate normalization of reads across our samples, at this stage of library construction we spiked-in small amounts of exogenous RNA from ArrayControlM-CM-"M-BM-^DM-BM-" Kit (Ambion) into each sample of poly(A)+ RNA. Paired-end sequencing was carried out by loading the samples onto four lanes (three samples per lane) of a flow cell and run on an Illumina Genome Analyzer IIx sequencer using 72 cycles per end of each paired-end read. Biological replicates of each genotype were loaded onto separate lanes.
Project description:These arrays measure gene expression across eight Y introgression lines in Drosophila simulans. Four lines (Ya19, Ya23, Ya24, Ya26) carry a D. simulans Y chromosome (from a Cameroon population) and four lines (Sec01, Sec03, Sec08, Sec27) carry a D. sechellia Y chromosome. All lines are otherwise identical with a D. simulans background (UCSD stock center line 14021-0251.092).
Project description:RNA was extracted from adult male and adult female Drosophila simulans carrying small genomic segments introgressed from Drosophila mauritiana Seven introgression genotypes were profiled, as well as the parental D. simulans and D. mauritiana strains. Each sex-by-genotype was assayed on four replicate arrays incorporating a dye swap
Project description:Males hybrids from the crosses between species of the D. simulans clade are steriles as the females are fertiles. Hybrid male sterility is due to severe defects in spermatogenesis and phenotypic differences are observed between the different hybrids involving D. simulans, D. sechellia and D. mauritiana. In this study we are comparing gene expression in the testes of hybrids involving the female D. simulans and the males D. melanogaster, D. mauritiana, or D. sechellia to the gene expression in species testes. Keywords: Comparative genomic hybridization 4 species (D. melanogaster, D. simulans, D. sechellia, D. mauritiana) and 3 different hybrids were used in this study. RNA extracted from whole D. melanogaster males was used as a reference. Hybridizations were perfomed using RNA extraxted from a pool of 200 testes from a sample with RNA extracted from whole D. melanogaster males. At least three independent replicates per hybridizations were performed
Project description:These arrays measure gene expression across eight Y introgression lines in Drosophila simulans. Four lines (Ya19, Ya23, Ya24, Ya26) carry a D. simulans Y chromosome (from a Cameroon population) and four lines (Sec01, Sec03, Sec08, Sec27) carry a D. sechellia Y chromosome. All lines are otherwise identical with a D. simulans background (UCSD stock center line 14021-0251.092). Four biological replicates of each of eight lines, plus two technical replicates (dye swap), for a total of 32 arrays. Full methods are described in the accompanying publication.
Project description:Males hybrids from the crosses between species of the D. simulans clade are steriles as the females are fertiles. Hybrid male sterility is due to severe defects in spermatogenesis and phenotypic differences are observed between the different hybrids involving D. simulans, D. sechellia and D. mauritiana. In this study we are comparing gene expression in the testes of hybrids involving the female D. simulans and the males D. melanogaster, D. mauritiana, or D. sechellia to the gene expression in species testes. Keywords: Comparative genomic hybridization
Project description:Species often produce sterile hybrids early in their evolutionary divergence, and some evidence suggests that hybrid sterility may be associated with deviations or disruptions in gene expression. In support of this idea, many studies have shown that a high proportion of male-biased genes are underexpressed compared to non-sex-biased genes in sterile F1 male hybrids of Drosophila species. In this study, we examined and compared patterns of misexpression in F1 hybrid male third instar larvae of Drosophila simulans and its sibling species, D. sechellia. We analyzed hybrids using custom cDNA arrays we developed from RT-PCRs of spermatogenesis-related transcripts from these species and another sibling species (D. mauritiana). The results from a commercial genome-wide array and custom chip for adults of this species pair, from the custom chip and the genome-wide chip for adults of the D. simulans-D. mauritiana species pair, and from the larvae of the D. simulans-D. mauritiana species pair, are presented separately. Keywords: Comparison of pure-species Drosophila expression to hybrid expression in larvae
Project description:Species often produce sterile hybrids early in their evolutionary divergence, and some evidence suggests that hybrid sterility may be associated with deviations or disruptions in gene expression. In support of this idea, many studies have shown that a high proportion of male-biased genes are underexpressed compared to non-sex-biased genes in sterile F1 male hybrids of Drosophila species. In this study, we examined and compared patterns of misexpression in F1 hybrid male third instar larvae of Drosophila simulans and its sibling species, D. mauritiana. We analyzed hybrids using custom cDNA arrays we developed from RT-PCRs of spermatogenesis-related transcripts from these species and another sibling species (D. sechellia). The results from a commercial genome-wide array and custom chip for adults of this species pair, from the custom chip and the genome-wide chip for adults of the D. simulans-D. sechellia species pair, and from the larvae of the D. simulans-D. sechellia species pair, are presented separately. Keywords: Comparison of pure-species Drosophila expression to hybrid expression