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Project 4: Isolation of organs and tissue

Isolation of organs and tissue from schistosomes


Recently, we established a novel method for the isolation, purification, and enrichment of whole intact reproductive organs of adult Schistosoma mansoni providing RNA as well as proteins of sufficient amounts as basis for tissue-specific analyses1. qRT-PCR analyses comparing transcript levels of selected genes based on RNA obtained from gonads and whole worms revealed a significant influence of the pairing status on the gene expression in ovary and testis, which was not traceable in whole worms2. More recently, the organ isolation protocol was adapted for isolating vitellaria and vitelline cells for further molecular and physiological characterisation3. Currently, comparative sub-transcriptomic RNA-seq analyses employing ovaries and testes from paired and unpaired adult schistosomes were performed revealing tissue-specific and paring-dependent gene expression patterns within the gonads of S. mansoni4,5. Results of these analyses will accelerate the elucidation of differentiation processes of the reproductive organs and the involved signal transduction processes. This will lead to putative molecular targets for novel concepts to combat schistosomiasis. Future plans aim at further developing and improving the novel method for the isolation of specific tissues from other Schistosoma life stages as well as investigating its applicability on other trematodes.

 

Ovare gepaarter WeibchenOvare (Eierstöcke) und Testes (Hoden) adulter Schistosomen (S. mansoni), die mit Hilfe der in unserem Labor etablierten Organ-Isolationsmethode gewonnen wurden1. Links: Ovare gepaarter Weibchen (sie enthalten mature, primäre Oocyten, dunkler Bereich, und unreife Oocyten, heller Bereich). Mitte: ein Ovar eines ungepaarten Weibchens (welches im Vergleich zu dem Ovar eines gepaarten Weibchens deutlich kleiner ist und ausschließlich unreife Oocyten aufweist). Rechts, Testes eines Schistosomen-Männchens.Testes

Ovaries and testes of S. mansoni obtained by the organ-isolation protocol, which was established in our lab1. Left, ovaries obtained from paired females (containing mature primary oocytes, dark part, and immature oocytes, light part). Middle, one ovary from an unpaired female (which compared to an ovary from a paired female is much smaller and contains only immature oocytes). Right, testes on a male schistosome.

 

1Hahnel S, Lu Z, Wilson RA, Grevelding CG, Quack T. (2013) Whole-organ isolation approach as a basis for tissue-specific analyses in Schistosoma mansoni. PLoS Negl Trop Dis. 7(7):e2336.

 

2Hahnel S, Quack T, Parker-Manuel SJ, Lu Z, Vanderstraete M, Morel M, Dissous C, Cailliau K, Grevelding CG. (2014) Gonad RNA-specific qRT-PCR analyses identify genes with potential functions in schistosome reproduction such as SmFz1 and SmFGFRs.

Front Genet. 5:170.

 

3Lu Z, Quack T, Hahnel S, Gelmedin V, Pouokam E, Diener M, Hardt M, Michel G, Baal N, Hackstein H, Grevelding CG. (2015) Isolation, enrichment and primary characterisation of vitelline cells from Schistosoma mansoni obtained by the organ isolation method. Int J Parasitol. 45(9-10):663-72.

 

4Lu Z, Sessler F, Holroyd N, Hahnel S, Quack T, Berriman M, Grevelding CG (2016) Gonad-specific and pairing-dependent gene expression in the blood fluke Schistosoma mansoni. Sci Rep. 6:31150.

 

5Lu, Z., Sessler, F., Holroyd, N., Hahnel, S., Quack, T., Berriman, M., Grevelding, C.G. (2017) A gene expression atlas of adult Schistosoma mansoni and their gonads. Scientific Data 4:170118.