Dataset Information

Acp36DE is required for uterine conformational changes in mated Drosophila females.

ABSTRACT: In a multitude of animals with internal fertilization, including insects and mammals, sperm are stored within a female's reproductive tract after mating. Defects in the process of sperm storage drastically reduce reproductive success. In Drosophila males, "Acp" seminal proteins alter female postmating physiology and behavior, and are necessary for several aspects of sperm storage. For example, Acps cause a series of conformational changes in the mated female's reproductive tract that occur during and immediately after mating. These conformational changes have been hypothesized to aid both in the movement of sperm within the female and in the subsequent storage of those sperm. We used RNAi to systematically knock down several Acps involved in sperm storage to determine whether they played a role in the mating-induced uterine conformational changes. Mates of males lacking the glycoprotein Acp36DE, which is needed for the accumulation of sperm in the storage organs, fail to complete the full sequence of the conformational changes. Our results show that uterine conformational changes are important for proper accumulation of sperm in storage and identify a seminal protein that mediates these changes. Four Acps included in this study, previously shown to affect sperm release from storage (CG9997, CG1656, CG1652, and CG17575), are not necessary for uterine conformational changes to occur. Rather, consistent with their role in later steps of sperm storage, we show here that their presence can affect the outcome of sperm competition situations.

SUBMITTER: Avila FW

PROVIDER: S-EPMC2747198 | biostudies-literature | 2009 Sep

REPOSITORIES: biostudies-literature

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Acp36DE is required for uterine conformational changes in mated Drosophila females.

Avila Frank W FW Wolfner Mariana F MF

Proceedings of the National Academy of Sciences of the United States of America 20090901 37

In a multitude of animals with internal fertilization, including insects and mammals, sperm are stored within a female's reproductive tract after mating. Defects in the process of sperm storage drastically reduce reproductive success. In Drosophila males, "Acp" seminal proteins alter female postmating physiology and behavior, and are necessary for several aspects of sperm storage. For example, Acps cause a series of conformational changes in the mated female's reproductive tract that occur durin ...[more]

PMID: 19805225

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Project description:In internally fertilizing organisms, mating involves a series of highly coordinated molecular interactions between the sexes that occur within the female reproductive tract. In species with promiscuous mating systems, traits involved in postcopulatory interactions are expected to evolve rapidly, potentially leading to postmating-prezygotic (PMPZ) reproductive isolation between diverging populations. Here, we use a novel study design to investigate the postmating transcriptional response of Drosophila mojavensis female reproductive tracts following copulation with either conspecific or heterospecific (D. arizonae) males at three time points postmating. Relatively few genes (15 total) were transcriptionally regulated in the female lower reproductive tract in response to conspecific mating. Heterospecifically-mated females exhibited significant perturbations in the expression of the majority of these genes, and also downregulated transcription of a number of others, including several involved in mitochondrial function. These striking regulatory differences indicate failed postcopulatory molecular interactions between the sexes consistent with the strong PMPZ isolation observed for this cross. We also report, for the first time, the transfer of male mRNA transcripts to females during copulation. These included transcripts from male accessory-gland proteins (ACPs), a finding with potentially broad implications for understanding postcopulatory molecular interactions between the sexes. Dataset from Postmating transcriptional changes in reproductive tracts of con- and heterospecifically-mated Drosophila mojavensis females Bono, JM, Matzkin,LM, Kelleher, ES, and Markow, MA, Proceedings of the National Academy of Sciences, USA. The treatments were con- and heterospecific mated D. mojavensis females. Three different post-copulation times points were assayed, 15 minutes, 2 hours and 6 hours. For each time point there were two independent replicates. Additionally, two replicates of a virgin D. mojavensis female control were assayed. A total of 14 arrays. All RNA extractions were from the lower reproductive tract of females.

Project description:Male-derived accessory gland proteins (Acps) that are transferred to females during mating have profound effects on female reproductive physiology including increased ovulation, mating inhibition, and effects on sperm utilization and storage. The extreme rates of evolution seen in Acps may be driven by sperm competition and sexual conflict, processes which may ultimately drive complex interactions between female- and male-derived molecules and sperm. However, little is known of how gene expression in female reproductive tissues changes in response to the presence of male molecules and sperm. To characterize this response, we conducted parallel genomic and proteomic analyses of gene expression in the reproductive tract of 3-day-old unmated and mated female Drosophila melanogaster. Using DNA microarrays, we identified 539 transcripts that are differentially expressed in unmated vs. mated females and revealed a striking peak in differential expression at 6 hrs postmating and a marked shift from primarily down-regulated to primarily up-regulated transcripts within 3 hrs after mating. Combining two-dimensional gel electrophoresis and liquid chromatography mass spectrometry analyses, we identified 84 differentially expressed proteins at 3 hrs postmating, including proteins which appeared to undergo post-translational modification. Together, our observations define transcriptional and translational response to mating within the female reproductive tract and suggest a bimodal model of postmating gene expression initially correlated with mating and the final stages of female reproductive tract maturation and later with the declining presence of male reproductive molecules and with sperm maintenance and utilization. Experiment Overall Design: Three-day-old mated and unmated females were dissected to remove the lower reproductive tract (upper uterus, sperm-storage organs, and accessory glands). Mated females were dissected either immediately following mating (0 hr) or at 3, 6, or 24 hrs following the termination of mating. Tracts of 12-40 females of like category were pooled and total RNA extracted via a TRIzol-based protocol. Processing and labeling of transcript was performed by the Molecular Biology Core Facility at the Medical College of Georgia. Arrays from mated females at the different timepoints were compared to unmated females.

Dataset Information

Acp36DE is required for uterine conformational changes in mated Drosophila females.

Publications

Acp36DE is required for uterine conformational changes in mated Drosophila females.

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