Project description:Fertility depends, in part, on interactions between male and female reproductive proteins inside the female reproductive tract (FRT) that mediate postmating changes in female behavior, morphology, and physiology. Coevolution between interacting proteins within species may drive reproductive incompatibilities between species, yet the mechanisms underlying postmating-prezygotic isolating barriers remain poorly resolved. Here, we used quantitative proteomics in sibling Drosophila species to investigate the molecular composition of the FRT environment and its role in mediating species-specific postmating responses. We found that (1) FRT proteomes in D. simulans and D. mauritiana virgin females express unique combinations of secreted proteins and are enriched for distinct functional categories, (2) mating induces substantial changes to the FRT proteome in D. mauritiana but not in D. simulans, and (3) the D. simulans FRT proteome exhibits limited postmating changes irrespective of whether females mate with conspecific or heterospecific males, suggesting an active female role in mediating reproductive interactions. Our study suggests that divergence in the FRT extracellular environment and postmating response contribute to previously described patterns of postmating-prezygotic isolation and the maintenance of species boundaries.
Project description:We sequenced dissected ovaries and testes (with reproductive tracts) as well as female and male carcasses in two species of Drosophila in order to validate gene predictions from the ModENCODE project. Comparison of dissected reproductive tracts and remaining carcasses between D. simulans and D. pseudoobscura
Project description:We compared 10, 20 and 40 animal pools to determine the best number of intact adults to pool Keywords: Agilent microarray, pooling, Drosophila, intact adults
Project description:MicroRNAs (miRNA) are small, endogenous RNAs that regulate the expression of mRNAs posttranscriptionally. Evolutionarily new miRNAs, like new protein-coding genes, are dominantly expressed in reproductive organs. To dissect the evolutionary dynamics of new miRNAs in Drosophila spp, we sequenced small RNAs from two species of Drosophila, including four samples from reproductive organs and one sample from imaginal discs / CNS. miRNA expression profile shows vast majority of new miRNAs are specifically expressed in testes and/or ovaries, suggesting a role of sexual selection for new miRNA evolution. Five small RNA samples, mainly from reproductive organs of D. simulans and D. pseudoobscura were analyzed. The small RNAs were sequenced by Illumina HiSeq 2000. After triming the adapters, the 18-30 nt sequences were extracted for further study.
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.