Transcriptional Analysis of a Unique Set of Genes Involved in Schistosoma mansoni Female Reproductive Biology

Abstract

Schistosomiasis affects more than 200 million people globally. The pathology of schistosome infections is due to chronic tissue inflammation and damage from immune generated granulomas surrounding parasite eggs trapped in host tissues. Schistosoma species are unique among trematode parasites because they are dioecious; females require paring with male parasites in order to attain reproductive maturity and produce viable eggs. Ex vivo cultured females lose the ability to produce viable eggs due to an involution of the vitellarium and loss of mature oocytes. In order to better understand schistosome reproductive biology we used data generated by serial analysis of gene expression (SAGE) to identify uncharacterized genes which have different transcript abundance in mature females, those that have been paired with males, and immature females obtained from unisexual infections. To characterize these genes we used bioinformatics, transcript localization, and transcriptional analysis during the regression of in vitro cultured females. Genes transcribed exclusively in mature females localize primarily in the vitellocytes and/or the ovary. Genes transcribed exclusively in females from single sex infections localize to vitellocytes and subtegumental cells. As female reproductive tissues regress, eggshell precursor proteins and genes involved in eggshell synthesis largely have decreased transcript abundance. However, some genes with elevated transcript abundance in mature adults have increased gene expression following regression indicating that the genes in this study function both in eggshell biology as well as vitellogenesis and maintenance of female reproductive tissues. In addition, we found that genes enriched in females from single sex infections have increased expression during regression in ex vivo females. By using these transcriptional analyses we can direct research to examine the areas of female biology that are both relevant to understanding the overall process of female development and worm pairing while determining novel therapeutic approaches directed at the maturation of female schistosomes.

Author Summary

Schistosomiasis is a chronic, debilitating disease that affects over 200 million people globally. The pathology associated with schistosomiasis is caused by host immune responses to parasite eggs. Therefore, it is imperative to identify pathways responsible for controlling worm reproductive biology. Schistosome females must be in constant contact with male parasites in order to achieve reproductive maturity. The process of pairing and reproductive maturation in female worms is poorly understood, in part, because it does not occur outside of the host. In addition, when female schistosomes are removed from their mammalian host they regress to an immature state. In this study our goal was to characterize a unique set of genes in Schistosoma mansoni whose transcript abundance differs in mature and immature female worms. We found that the genes with higher transcript abundance in sexually mature female worms were expressed in female reproductive tissues, while those transcripts enriched in sexually immature worms were present in sub-surface somatic cells. Transcript abundance of the selected genes changed dramatically when females were removed from their host. These findings inform new approaches to study female worm biology and will provide insights into the processes of worm pairing and reproductive maturation.