Post-mating gene expression of Mexican fruit fly females: disentangling the effects of the male accessory glands.

Mating has profound physiological and behavioural consequences for female insects. During copulation, female insects typically receive not only sperm, but a complex ejaculate containing hundreds of proteins and other molecules from male reproductive tissues, primarily the reproductive accessory glands. The post-mating phenotypes affected by male accessory gland (MAG) proteins include egg development, attraction to oviposition hosts, mating, attractiveness, sperm storage, feeding and lifespan. In the Mexican fruit fly, Anastrepha ludens, mating increases egg production and the latency to remating. However, previous studies have not found a clear relationship between injection of MAG products and oviposition or remating inhibition in this species. We used RNA-seq to study gene expression in mated, unmated and MAG-injected females to understand the potential mating- and MAG-regulated genes and pathways in A. ludens. Both mating and MAG-injection regulated transcripts and pathways related to egg development. Other transcripts regulated by mating included those with orthologs predicted to be involved in immune response, musculature and chemosensory perception, whereas those regulated by MAG-injection were predicted to be involved in translational control, sugar regulation, diet detoxification and lifespan determination. These results suggest new phenotypes that may be influenced by seminal fluid molecules in A. ludens. Understanding these influences is critical for developing novel tools to manage A. ludens.

The consequences of genetic variation in sex peptide expression levels for egg laying and retention in females.

The accessory gland proteins (Acps) that male Drosophila melanogaster produce and transfer to females during copulation are key to male and female fitness. One Acp, the sex peptide (SP), is largely responsible for a dramatic increase in female egg laying and decrease in female receptivity after copulation. While genetic variation in male SP expression levels correlate with refractory period duration in females, it is unknown whether male SP expression influences female egg laying or if any effect of SP is mediated by SP retention in the female reproductive tract. Here we measured the amount of SP retained in the female reproductive tract after mating and female egg laying after copulating with virgin males. We found no correlation between male SP expression levels and egg laying, or the amount of SP in the female reproductive tract after mating. Additionally, the amount of SP retained in the female did not influence egg laying. These finding suggests that additional factors, such as variation in other Acps, are important for the retention of SP in females and its quantitative effects on egg laying. It also shows that egg laying and refractory period response to SP is at least partially uncoupled.

Identification of predicted seminal fluid proteins in Tribolium castaneum.

In several insect species, seminal fluid proteins (SFPs) have been demonstrated to be key regulators of male and female fitness through their ability to alter female physiology and behaviour. Tribolium castaneum is an economically important pest species and a model system for sexual selection research, but little is known about SFPs in this insect. To create a foundation for the study of T. castaneum SFPs, we used mass spectrometry to identify putative SFPs by comparing proteins detected in the male reproductive glands with those found in the reproductive tracts of virgin and mated females. Fourteen putative SFPs, thirteen with male biased expression, were identified through this approach. We also used reverse transcription PCR (RT-PCR) to examine expression levels across different tissue types. We found strongly male-biased expression in 13 genes, nine of which were expressed only in male accessory gland tissue. This represents the first proteomic-based method of identifying putative SFPs in any coleopteran species, and is the first study in this species to identify putative SFPs that are likely transferred to the female. This work could lead to functional analyses of the role of SFPs in sexual selection, sexual conflict and potential control of a pest species.