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New insights into the reproductive transcriptome profile of the Mediterranean fruit fly Ceratitis capitata

ABSTRACT: Purpose: Use of RNAseq to identify crucial genes expressed in the reproductive tract of C. capitata The medfly, Ceratitis capitata continues to pose a major threat to the agricultural sector in almost every continent. Promiscuous female medflies can oviposit up to 1000 eggs throughout their lifetime causing remarkable losses in crop viability. Nowadays, there is an increasing demand for finding new improved solutions to counteract its invasive spreading capacity. To these regards, research that aims in understanding the reproductive biology of fruit fly pests by obtaining its genomic profiling can be of great contribution to this field. In this study we have generated a De-novo assembly of the medfly reproductive tract transcripts. We used Transcriptomics via deep RNA-sequencing to identify the transcripts present in the sex organs (spermatheca, uterus and accessory glands) of sexually matured virgin and 24hr-freshly mated females as well as testes, and male accessory glands of C. capitata. In addition, differential expression changes between virgin and mated females could help elucidate crucial candidate genes that participate in the female physiological response to the male seminal input. Furthermore, in order to ensure pure and significant results of the transcriptomic analysis, we have included a negative control that includes gut and abdominal adipose tissue. The newly annotated genes from this investigation could furthermore contribute to the ongoing field of biological and genetic population control.

ORGANISM(S): Ceratitis capitata

PROVIDER: GSE80605 | GEO | 2016/04/26

SECONDARY ACCESSION(S): PRJNA319480

REPOSITORIES: GEO

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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.

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New insights into the reproductive transcriptome profile of the Mediterranean fruit fly Ceratitis capitata

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