- Background: Despite the growing interest in the female side of copulatory interactions, the roles played by differential expression and alternative splicing mechanisms of pre-RNA on tissues outside of the reproductive tract have remained largely unknown. - Here we addressed these questions in the context of con- vs heterospecific matings between Drosophila mojavensis and its sister species, D. - To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.. - The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.. - The role of seminal fluid proteins in the female postmating response has been well character- ized in Drosophila species, and hundreds of male- derived proteins have now been identified in other taxa [4–7]. - Perturbation of the female transcriptional response by heterospecific matings was first demonstrated in female reproductive tracts of Drosophila mojavensis when mated to D. - It defines the strength of the con- vs heterospecific response (Fig. - 3) as well as when such changes are induced in the heads (45 min vs 6 h postmating periods). - The number of genes and the distribution of the expression response in DE genes were substantially perturbed by the heterospecific matings (Fig. - The re- sponse of ♀Dmoj was stronger for conspecific matings in terms of the number of genes involved (Fig. - ♀Dari involved more genes in the heterospecific matings. - than that of the conspecific matings (Fig. - How- ever, a few of these genes show interesting and opposing patterns between con- and heterospecific matings, which make them additionally interesting candidates to further investigate in the context of the reproductive isolation. - 4a), which showed a moderate correlation in the conspecific-mating response between the species (♀Dmoj con vs ♀Dari con, Spearman’s ρ = 0.51), the rest of the comparisons (DE and AS) were not correlated be- tween the species (con- or heterospecific) (Fig. - 0.05) following GLM analysis are indicated with * in the “ down ” plot. - Secondly, DE genes exhibited substantial differences in the evolutionary rates between both species and crosses. - Pathways associated with i) nutrient homeostasis are particularly interesting for their implications in the female postmating response.. - Error bars represent standard error of the mean. - Most of the enriched pathways were detected in a conspecific context, but only a subset remained significant in the heterospecific matings (Fig. - Our results indicate that mating induces not only gene expression changes in female heads soon after mating, but also that a great part of the female postmating re- sponse involves a change in alternative splicing. - The number of genes responding through AS often exceeded that of DE genes, but both mechanisms appear to be in- volved in the female postcopulatory response. - Most of these are related to earlier post- mating processes and molecular interactions occurring in the female tract [e.g. - In this study, the expression of proteolytic cascades we observed in heads of mated females does not seem connected to functions occurring in the female reproductive tract. - Given the distance and heterogeneity of the tissues involved, changes induced in the female head are assumed to be the product of altered interaction networks, influencing the physiology of other tissues [29, 60]. - expression at distant sites in the fly body [61]. - Further research is needed to disentangle the underlying mechanisms causing the activation of genes outside of the female reproductive tract and their involvement in the post-copulatory response.. - Although female reproductive tract genes involved in the postmating response have only been investigated in a few species, results from D. - melanogaster, where courtship is longer and involves visual displays in front of the female [65, 66], courtship in cactophilic Drosoph- ila likely involves chemical and auditory cues, as it oc- curs largely behind the female [64]. - 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