scholarly journals The Drosophila melanogaster Seminal Fluid Protein Acp62F Is a Protease Inhibitor That Is Toxic Upon Ectopic Expression

Genetics ◽  
2002 ◽  
Vol 160 (1) ◽  
pp. 211-224
Author(s):  
Oliver Lung ◽  
Uyen Tram ◽  
Casey M Finnerty ◽  
Marcie A Eipper-Mains ◽  
John M Kalb ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Protease Inhibitor ◽  
Life Span ◽  
Reproductive Tract ◽  
Seminal Fluid ◽  
Ectopic Expression ◽  
Circulatory System ◽  
Sperm Storage ◽  
Egg Laying ◽  
Seminal Fluid Proteins

Abstract Drosophila melanogaster seminal fluid proteins stimulate sperm storage and egg laying in the mated female but also cause a reduction in her life span. We report here that of eight Drosophila seminal fluid proteins (Acps) and one non-Acp tested, only Acp62F is toxic when ectopically expressed. Toxicity to preadult male or female Drosophila occurs upon one exposure, whereas multiple exposures are needed for toxicity to adult female flies. Of the Acp62F received by females during mating, ~10% enters the circulatory system while ~90% remains in the reproductive tract. We show that in the reproductive tract, Acp62F localizes to the lumen of the uterus and the female's sperm storage organs. Analysis of Acp62F's sequence, and biochemical assays, reveals that it encodes a trypsin inhibitor with sequence and structural similarities to extracellular serine protease inhibitors from the nematode Ascaris. In light of previous results demonstrating entry of Acp62F into the mated female's hemolymph, we propose that Acp62F is a candidate for a molecule to contribute to the Acp-dependent decrease in female life span. We propose that Acp62F's protease inhibitor activity exerts positive protective functions in the mated female's reproductive tract but that entry of a small amount of this protein into the female's hemolymph could contribute to the cost of mating.

scholarly journals Mated Drosophila melanogaster Females Require a Seminal Fluid Protein, Acp36DE, to Store Sperm Efficiently

Genetics ◽  
1999 ◽  
Vol 153 (2) ◽  
pp. 845-857 ◽  
Author(s):  
Deborah M Neubaum ◽  
Mariana F Wolfner
Keyword(s):  
Drosophila Melanogaster ◽  
Seminal Fluid ◽  
Sperm Storage ◽  
Egg Laying ◽  
Limited Area ◽  
Seminal Fluid Proteins ◽  
Barrier Formation ◽  
Storage Organs ◽  
Initial Storage ◽  
Seminal Fluid Protein

Abstract Mated females of many animal species store sperm. Sperm storage profoundly influences the number, timing, and paternity of the female’s progeny. To investigate mechanisms for sperm storage in Drosophila melanogaster, we generated and analyzed mutations in Acp36DE. Acp36DE is a male seminal fluid protein whose localization in mated females suggested a role in sperm storage. We report that male-derived Acp36DE is essential for efficient sperm storage by females. Acp36DE1 (null) mutant males produced and transferred normal amounts of sperm and seminal fluid proteins. However, mates of Acp36DE1 males stored only 15% as many sperm and produced 10% as many adult progeny as control-mated females. Moreover, without Acp36DE, mated females failed to maintain an elevated egg-laying rate and decreased receptivity, behaviors whose persistence (but not initiation) normally depends on the presence of stored sperm. Previous studies suggested that a barrier in the oviduct confines sperm and Acp36DE to a limited area near the storage organs. We show that Acp36DE is not required for barrier formation, but both Acp36DE and the barrier are required for maximal sperm storage. Acp36DE associates tightly with sperm. Our results indicate that Acp36DE is essential for the initial storage of sperm, and that it may also influence the arrangement and retention of stored sperm.

scholarly journals Male Seminal Fluid Proteins Are Essential for Sperm Storage in Drosophila melanogaster

Genetics ◽  
1999 ◽  
Vol 153 (2) ◽  
pp. 837-844 ◽  
Author(s):  
Uyen Tram ◽  
Mariana F Wolfner
Keyword(s):  
Drosophila Melanogaster ◽  
Seminal Fluid ◽  
Accessory Gland ◽  
Sperm Storage ◽  
Competitive Pcr ◽  
Seminal Fluid Proteins ◽  
Pcr Assays ◽  
Transgenic Drosophila ◽  
Direct Counts

AbstractThe seminal fluid that is transferred along with sperm during mating acts in many ways to maximize a male’s reproductive success. Here, we use transgenic Drosophila melanogaster males deficient in the seminal fluid proteins derived from the accessory gland (Acps) to investigate the role of these proteins in the fate of sperm transferred to females during mating. Competitive PCR assays were used to show that while Acps contribute to the efficiency of sperm transfer, they are not essential for the transfer of sperm to the female. In contrast, we found that Acps are essential for storage of sperm by females. Direct counts of stored sperm showed that 10% of normal levels are stored by females whose mates transfer little or no Acps along with sperm.

scholarly journals Quantitative proteomics identification of seminal fluid proteins in male Drosophila melanogaster

2018 ◽  
Author(s):  
Irem Sepil ◽  
Ben R Hopkins ◽  
Rebecca Dean ◽  
Marie-Laëtitia Thézénas ◽  
Philip D Charles ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Quantitative Proteomics ◽  
Reproductive Tract ◽  
Seminal Fluid ◽  
High Sensitivity ◽  
Female Reproductive Tract ◽  
Label Free ◽  
Seminal Fluid Proteins ◽  
Copy Numbers ◽  
Before And After

AbstractSeminal fluid contains some of the fastest evolving proteins currently known. These seminal fluid proteins (Sfps) play crucial roles in reproduction, such as supporting sperm function, and – particularly in insects – modifying female physiology and behaviour. Identification of Sfps in small animals is challenging, and often relies on samples taken from the female reproductive tract after mating. A key pitfall of this method is that it might miss Sfps that are of low abundance due to dilution in the female-derived sample or rapid processing in females. Here we present a new and complimentary method, which provides added sensitivity to Sfp identification. We applied label-free quantitative proteomics to Drosophila melanogaster male reproductive tissue – where Sfps are unprocessed, and highly abundant – and quantified Sfps before and immediately after mating, to infer those transferred during copulation. We also analysed female reproductive tracts immediately before and after copulation to confirm the presence and abundance of known and candidate Sfps, where possible. Results were cross-referenced with transcriptomic and sequence databases to improve confidence in Sfp detection. Our data was consistent with 124 previously reported Sfps. We found 8 high-confidence novel candidate Sfps, which were both depleted in mated versus unmated males and identified within the reproductive tract of mated but not virgin females. We also identified 31 more candidates that are likely Sfps based on their abundance, known expression and predicted characteristics, and revealed that four proteins previously identified as Sfps are at best minor contributors to the ejaculate. The estimated copy numbers for our candidate Sfps were lower than for previously identified Sfps, supporting the idea that our technique provides a deeper analysis of the Sfp proteome than previous studies. Our results demonstrate a novel, high-sensitivity approach to the analysis of seminal fluid proteomes, whose application will further our understanding of reproductive biology.

scholarly journals Differences in Postmating Transcriptional Responses between Conspecific and Heterospecific Matings in Drosophila

2020 ◽  
Author(s):  
Yasir H Ahmed-Braimah ◽  
Mariana F Wolfner ◽  
Andrew G Clark
Keyword(s):  
Reproductive Tract ◽  
Evolutionary Dynamics ◽  
Seminal Fluid ◽  
Female Reproductive Tract ◽  
Additional Stress ◽  
Reproductive Proteins ◽  
Transcriptional Responses ◽  
Seminal Fluid Proteins ◽  
Reproductive Genes ◽  
Imd Pathway

Abstract In many animal species, females undergo physiological and behavioral changes after mating. Some of these changes are driven by male-derived seminal fluid proteins and are critical for fertilization success. Unfortunately, our understanding of the molecular interplay between female and male reproductive proteins remains inadequate. Here, we analyze the postmating response in a Drosophila species that has evolved strong gametic incompatibility with its sister species; Drosophila novamexicana females produce only ∼1% fertilized eggs in crosses with Drosophila americana males, compared to ∼98% produced in within-species crosses. This incompatibility is likely caused by mismatched male and female reproductive molecules. In this study, we use short-read RNA sequencing to examine the evolutionary dynamics of female reproductive genes and the postmating transcriptome response in crosses within and between species. First, we found that most female reproductive tract genes are slow-evolving compared to the genome average. Second, postmating responses in con- and heterospecific matings are largely congruent, but heterospecific matings induce expression of additional stress-response genes. Some of those are immunity genes that are activated by the Imd pathway. We also identify several genes in the JAK/STAT signaling pathway that are induced in heterospecific, but not conspecific mating. While this immune response was most pronounced in the female reproductive tract, we also detect it in the female head and ovaries. These results show that the female’s postmating transcriptome-level response is determined in part by the genotype of the male, and that divergence in male reproductive genes and/or traits can have immunogenic effects on females.

scholarly journals Sex peptide of Drosophila melanogaster males is a global regulator of reproductive processes in females

2012 ◽  
Vol 279 (1746) ◽  
pp. 4423-4432 ◽  
Author(s):  
A. Gioti ◽  
S. Wigby ◽  
B. Wertheim ◽  
E. Schuster ◽  
P. Martinez ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Sexual Conflict ◽  
Seminal Fluid ◽  
Nutrient Sensing ◽  
Seminal Fluid Proteins ◽  
Sex Peptide ◽  
Efficient Induction ◽  
Transcriptional Changes ◽  
Female Behaviour ◽  
Reproductive Processes

Seminal fluid proteins (Sfps) alter female behaviour and physiology and can mediate sexual conflict. In Drosophila melanogaster , a single Sfp, the sex peptide (SP), triggers remarkable post-mating responses in females, including altered fecundity, feeding, immunity and sexual receptivity. These effects can favour the evolutionary interests of males while generating costs in females. We tested the hypothesis that SP is an upstream master-regulator able to induce diverse phenotypes through efficient induction of widespread transcriptional changes in females. We profiled mRNA responses to SP in adult female abdomen (Abd) and head+thorax (HT) tissues using microarrays at 3 and 6 h following mating. SP elicited a rich, subtle signature of temporally and spatially controlled mRNAs. There were significant alterations to genes linked to egg development, early embryogenesis, immunity, nutrient sensing, behaviour and, unexpectedly, phototransduction. There was substantially more variation in the direction of differential expression across time points in the HT versus Abd. The results support the idea that SP is an important regulator of gene expression in females. The expression of many genes in one sex can therefore be under the influence of a regulator expressed in the other. This could influence the extent of sexual conflict both within and between loci.

scholarly journals Differences in post-mating transcriptional responses between conspecific and heterospecific matings in Drosophila

2020 ◽  
Author(s):  
Yasir H. Ahmed-Braimah ◽  
Mariana F. Wolfner ◽  
Andrew G. Clark
Keyword(s):  
Reproductive Tract ◽  
Evolutionary Dynamics ◽  
Seminal Fluid ◽  
Female Reproductive Tract ◽  
Additional Stress ◽  
Reproductive Proteins ◽  
Transcriptional Responses ◽  
Seminal Fluid Proteins ◽  
Reproductive Genes ◽  
Imd Pathway

AbstractIn many animal species, females undergo physiological and behavioral changes after mating. Some of these changes are driven by male-derived seminal fluid proteins, and are critical for fertilization success. Unfortunately, our understanding of the molecular interplay between female and male reproductive proteins remains superficial. Here we analyze the post-mating response in a Drosophila species that has evolved strong gametic incompatibility with its sister species; D. novamexicana females produce only 1% fertilized eggs in crosses with D. americana males, compared to ~98% produced in within-species crosses. This incompatibility is likely caused by mismatched male and female reproductive molecules. In this study we use short-read RNA sequencing to examine the evolutionary dynamics of female reproductive genes and the post-mating transcriptome response in crosses within and between species. First, we found that most female reproductive tract genes are slow-evolving compared to the genome average. Second, post-mating responses in con- and heterospecific matings are largely congruent, but heterospecific mating induces expression of additional stress-response genes. Some of those are immunity genes that are activated by the Imd pathway. We also identify several genes in the JAK/STAT signaling pathway that are induced in heterospecific, but not conspecific mating. While this immune response was most pronounced in the female reproductive tract, we also detect it in the female head and ovaries. Our results show that the female’s post-mating transcriptome-level response is determined in part by the genotype of the male, and that divergence in male reproductive genes and/or traits can have immunogenic effects on females.

scholarly journals Rapid divergence of the copulation proteins in the Drosophila dunni group is associated with hybrid post-mating-prezygotic incompatibilities

2020 ◽  
Author(s):  
Tom Hill ◽  
Hazel-Lynn Rosales-Stephens ◽  
Robert L. Unckless
Keyword(s):  
Seminal Fluid ◽  
Rapid Evolution ◽  
Species Group ◽  
Egg Laying ◽  
Reproductive Proteins ◽  
Seminal Fluid Proteins ◽  
Prezygotic Isolation ◽  
Caribbean Islands ◽  
Female Survival ◽  
Rapid Divergence

AbstractProteins involved in post-copulatory interactions between males and females are among the fastest evolving genes in many species and this has been attributed to reproductive conflict. Likely as a result, these proteins are frequently involved in cases of post-mating-prezygotic isolation between species. The Drosophila dunni subgroup consists of a dozen recently diverged species found across the Caribbean islands with varying levels of hybrid incompatibility. We sought to examine how post-mating-prezygotic factors are involved in isolation among members of this species group. We performed experimental crosses between species in the dunni group and find evidence of hybrid inviability. We also find an insemination reaction-like response preventing egg laying and leading to reduced female survival post-mating. To identify that genes may be involved in these incompatibilities, we sequenced and assembled the genomes of four species in the dunni subgroup and looked for signals of rapid evolution between species. Despite low levels of divergence, we found evidence of rapid evolution and divergence of some reproductive proteins, specifically the seminal fluid proteins. This suggests post-mating-prezygotic isolation as a barrier for gene flow between even the most closely related species in this group and seminal fluid proteins as a possible culprit.

scholarly journals Interactions between the microbiome and mating influence the female’s transcriptional profile in Drosophila melanogaster

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Sofie Y. N. Delbare ◽  
Yasir H. Ahmed-Braimah ◽  
Mariana F. Wolfner ◽  
Andrew G. Clark
Keyword(s):  
Drosophila Melanogaster ◽  
Egg Production ◽  
Seminal Fluid ◽  
Transcript Abundance ◽  
Vital Role ◽  
Transcriptional Profile ◽  
Seminal Fluid Proteins ◽  
Two Factors ◽  
Immunity Genes ◽  
Neuronal Functions

Abstract Drosophila melanogaster females undergo a variety of post-mating changes that influence their activity, feeding behavior, metabolism, egg production and gene expression. These changes are induced either by mating itself or by sperm or seminal fluid proteins. In addition, studies have shown that axenic females—those lacking a microbiome—have altered fecundity compared to females with a microbiome, and that the microbiome of the female’s mate can influence reproductive success. However, the extent to which post-mating changes in transcript abundance are affected by microbiome state is not well-characterized. Here we investigated fecundity and the post-mating transcript abundance profile of axenic or control females after mating with either axenic or control males. We observed interactions between the female’s microbiome and her mating status: transcripts of genes involved in reproduction and genes with neuronal functions were differentially abundant depending on the females’ microbiome status, but only in mated females. In addition, immunity genes showed varied responses to either the microbiome, mating, or a combination of those two factors. We further observed that the male’s microbiome status influences the fecundity of both control and axenic females, while only influencing the transcriptional profile of axenic females. Our results indicate that the microbiome plays a vital role in the post-mating switch of the female’s transcriptome.

A genome-wide analysis of courting and mating responses in Drosophila melanogaster females

Genome ◽  
2004 ◽  
Vol 47 (5) ◽  
pp. 900-910 ◽  
Author(s):  
Mara KN Lawniczak ◽  
David J Begun
Keyword(s):  
Gene Expression ◽  
Drosophila Melanogaster ◽  
Seminal Fluid ◽  
Differentially Expressed ◽  
Male Reproductive Success ◽  
Female Side ◽  
Accessory Gland Proteins ◽  
Seminal Fluid Proteins ◽  
A Genome ◽  
Immune Related Genes

In Drosophila melanogaster, seminal fluid proteins influence several components of female physiology and behavior, including re-mating rates, ovulation and oviposition, and sperm use. It is well-known that female flies are not simply passive vessels and that female-mediated interactions with male products are important to female (and thus male) reproductive success. While the population genetics, molecular evolution and physiological effects of seminal fluid proteins have been examined, the genetics and evolution of the female side of these post-mating interactions is unexplored in spite of work showing that female genotype and female-by-male genotype interactions are important determinants of sperm competition outcomes. Here we use microarrays to identify candidate genes involved in the female side of post-mating sexual interactions. We report the results of a whole-genome oligonucleotide chip experiment that reveals 23 genes differentially expressed between virgin females exposed and unexposed to courting males, and 38 genes differentially expressed between virgin and recently mated females. Immune related genes are overrepresented among the mating-influenced candidates. We use quantitative reverse-transcriptase PCR to independently assess gene expression changes for roughly half of the mating-affected candidate genes.Key words: reproduction, gene expression, Drosophila immune related genes, serine proteases, accessory gland proteins.

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