Project 2: Characterisation of mitosis and egg production associated genes
Characterisation of mitosis- and egg production-associated genes in Schistosoma mansoni
Since the eggs are essential for the parasites lifecycle and responsible for the pathology of schistosomiasis1, genes which are involved in the reproduction of S. mansoni are studied in this project in detail. Basis for this project were results of in vitro experiments with inhibitors against TGFβ- and the Src-kinase pathways. Using these inhibitors showed effects on pairing stability and the egg production of adult schistosomes in vitro. Subsequent microarray analyses with RNA from inhibitor-treated worms revealed many differential expressed genes. Among these were Ca2+ metabolism-associated genes and further genes which are probably involved in transcriptional processes2.
In order to characterize these genes a set of methods will be used (investigation of tissue-specific gene transcription via in situ-hybridization and organ-specific RT-PCRs/qPCR analyses; yeast two-hybrid interaction studies to identify binding partners and to unravel signal transduction processes; Ca2+-imaging-based cell assays to examine the influence of inhibitors against Ca2+-homeostasis; RNAi-and/or inhibitor experiments followed by the analyses of physiological and morphological effects). Aim of this study is to gain more insights into the molecular basics of pairing-dependent differentiation of reproduction-organs in schistosomes.
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Figure A: Microscopic analyses (confocal laser scanning micrsocopy3) of schistosome couples treated with an inhibitor against a protein involved in Ca2+-homeostasis. After 7 days of treatment dark areas appeared inside the female vitellarium (vi) in comparison to untreated controls (B). These perforated areas give a first hint towards a potential loss of tissue integrity inside the vitellarium. 1 = male, 2 = vitellarium of the female, 3 = gut of the female. |
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1WHO factsheets Schistosomiasis (2015) http://www.who.int/mediacentre/factsheets/fs115/en/
2Buro C, Oliveira KC, Lu Z, Leutner S, Beckmann S, Dissous C, Cailliau K, Verjovski-Almeida S, Grevelding CG. (2013) Transcriptome analyses of inhibitor-treated schistosome females provide evidence for cooperating Src-kinase and TGFß receptor pathways controlling mitosis and eggshell formation. PLOS Pathogens 9(6):e1003448.
3 Beckmann S, Quack T, Burmeister C, Buro C, Long T, Dissous C, Grevelding CG. (2010)
Schistosoma mansoni: signal transduction processes during the development of the reproductive organs. Parasitology 137(3):497-520.