Project description:Fertility depends on the coordination of complex interactions between male and female reproductive proteins inside the female reproductive tract (FRT). These interactions mediate a suite of changes in female behavior, morphology, and physiology after mating, yet little is known about how the molecular environment of the FRT may differ among species and coordinate species-specific female post-mating responses. We used semi-quantitative proteomics to compare the FRT protein composition between virgin and mated females in Drosophila melanogaster. These results are compared to those from quantitative TMT proteomic analyses of the mating-induced changes in D. simulans and D. mauritiana, and after conspecific and heterospecific inseminations. Our study highlights the value of using quantitative proteomics approaches to study the molecular composition of the FRT environment, and how its divergence may inform mechanistic studies of post-mating pre-zygotic reproductive isolation between species.
Project description:Comparison of females mated to males null for Sex-Peptide (SP0, Liu, H. and E. Kubli, Sex-peptide is the molecular basis of the sperm effect in Drosophila melanogaster. Proc Natl Acad Sci U S A, 2003. 100(17): p. 9929-33) or to control, Sex-Peptide producing, males. Comparisons were made at 3 and 6 hours after mating, in dissected Head-Thorax body parts.
Project description:Comparison of females mated to males null for Sex-Peptide (SP0, Liu, H. and E. Kubli, Sex-peptide is the molecular basis of the sperm effect in Drosophila melanogaster. Proc Natl Acad Sci U S A, 2003. 100(17): p. 9929-33) or to control, Sex-Peptide producing, males. Comparisons were made at 3 and 6 hours after mating, in dissected Abdomen parts.<br>
Project description:Comparison of females mated to males null for Sex-Peptide (SP0, Liu, H. and E. Kubli, Sex-peptide is the molecular basis of the sperm effect in Drosophila melanogaster. Proc Natl Acad Sci U S A, 2003. 100(17): p. 9929-33) or to control, Sex-Peptide producing, males. Comparisons were made at 3 and 6 hours after mating, in in dissected Abdomen body parts.
Project description:Comparison of females mated to males null for Sex-Peptide (SP0, Liu, H. and E. Kubli, Sex-peptide is the molecular basis of the sperm effect in Drosophila melanogaster. Proc Natl Acad Sci U S A, 2003. 100(17): p. 9929-33) or to control, Sex-Peptide producing, males. Comparisons were made at 3 and 6 hours after mating, in dissected Head - Thorax body parts.
Project description:Sex Peptide, a seminal fluid protein of Drosophila melanogaster males, elicits an array of post-mating responses in females, including decreased receptivity to re-mating, and increased egg laying, activity and food intake, with a preference for protein-rich food. To determine how one protein can have such widespread effects, we set out to dissect the genetic architecture of the female’s response to Sex Peptide, to determine whether Sex Peptide alters the expression of several regulators targeted to specific post-mating responses or acts on a pleiotropic regulator that controls multiple responses. We performed bulk RNA-seq of female heads at 10 time points within the first 24 hours after mating, sampling virgin females, females mated to control males and females mated to Sex Peptide-null males. Using this high-resolution time series, we identified mating- and Sex Peptide-dependent differentially expressed genes and discovered the presence of differentially used exons. We constructed gene regulatory networks using clustering and motif enrichment analyses, and identified cell types in which these changes might take place using deconvolution of our bulk RNA-seq dataset. One key network included metabolic genes which might change in expression in the female’s fat body. A second network included genes with neuronal functions, whose changes might be located in neurons or sensory organs in the female’s head. Within these networks we identified known molecular regulators of the circadian clock. Further, we found that many differentially expressed genes, and some differentially used exons, followed a circadian rhythm in virgin females, and that this rhythm was altered after mating with a Sex Peptide- male.
Project description:Drosophila adult midgut genes with sex-biased transcription and/or splicing Sex differences in physiology are commonly attributed to developmental and/or hormonal factors, but there is increasing realisation that cell-intrinsic mechanisms play important and persistent roles. Here we use the Drosophila melanogaster intestine to investigate the activity and significance of intrinsic sex in an adult somatic organ in vivo. We find that the adult intestinal epithelium is a cellular mosaic of different sex differentiation pathways, and displays extensive sex differences in expression of genes with roles in growth and metabolism. Cell-specific reversals of the sexual identity of adult intestinal stem cells uncover its key roles in controlling organ size, its reproductive plasticity and susceptibility to tumours. Unlike previous examples of sexually dimorphic somatic stem cell activity, the sex differences in intestinal stem cell behaviour arise from intrinsic mechanisms, which control cell cycle duration and involve a new doublesex- and fruitless-independent branch of the sex differentiation pathway downstream of transformer. Together, our findings indicate that the plasticity of an adult somatic organ is reversibly controlled by its intrinsic sexual identity, imparted by a new mechanism that may be active in more tissues than previously recognised. Adult midgut transcriptomes of 15-day-old virgin female and males were generated by deep sequencing, in triplicate using a Hiseq2000 using paired end 100bp reads.
Project description:Methionine restriction is known to extend lifespan in various model organisms including Drosophila melanogaster. In this analysis, we performed scRNAseq of Drosophila female midgut samples to understand the cell type specific response to methionine restriction.
Project description:Identification of midgut genes with sex-biased transcription and/or splicing under cell-autonomous control of transformer in adult intestinal stem cells Sex differences in physiology are commonly attributed to developmental and/or hormonal factors, but there is increasing realisation that cell-intrinsic mechanisms play important and persistent roles. Here we use the Drosophila melanogaster intestine to investigate the activity and significance of intrinsic sex in an adult somatic organ in vivo. We find that the adult intestinal epithelium is a cellular mosaic of different sex differentiation pathways, and displays extensive sex differences in expression of genes with roles in growth and metabolism. Cell-specific reversals of the sexual identity of adult intestinal stem cells uncover its key roles in controlling organ size, its reproductive plasticity and susceptibility to tumours. Unlike previous examples of sexually dimorphic somatic stem cell activity, the sex differences in intestinal stem cell behaviour arise from intrinsic mechanisms, which control cell cycle duration and involve a new doublesex- and fruitless-independent branch of the sex differentiation pathway downstream of transformer. Together, our findings indicate that the plasticity of an adult somatic organ is reversibly controlled by its intrinsic sexual identity, imparted by a new mechanism that may be active in more tissues than previously recognised. Adult midgut transcriptomes of 15-day-old virgin females were generated by deep sequencing, in triplicate using a Hiseq2000 using paired end 100bp reads. Genotypes were: control, transformer mutant and transformer mutant female in which transformer was re-introduced in adult intestinal stem cells.