Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Functional genomic characterization of germinal cells in larval Schistosoma mansoni

ABSTRACT: RNA-seq experiments to identify gene expression changes following miracidia to sporocyst transformation in Schistosoma mansoni. Schistosomes infect hundreds of millions of people in the developing world. Transmission of these parasites relies on a stem cell-driven, clonal expansion of larvae inside a molluscan intermediate host. How this novel asexual reproductive strategy relates to current models of stem cell maintenance and germline specification is unclear. Here, we demonstrate that this proliferative larval cell population (germinal cells) shares some molecular signatures with stem cells from diverse organisms, in particular neoblasts of planarians (free-living relatives of schistosomes). We identify two distinct germinal cell lineages that differ in their proliferation kinetics and expression of a nanos ortholog. We show that a vasa PL10 homolog is required for proliferation and maintenance of both populations; whereas, argonaute2 and a fibroblast growth factor receptor-encoding gene are required only for nanos-negative cells. Our results suggest that an ancient stem cell-based developmental program may have enabled the evolution of the complex life-cycle of parasitic flatworms. Total RNA was purified from ~10,000 freshly hatched miracidia or sporocysts 48 h post-transformation.

ORGANISM(S): Schistosoma mansoni

SUBMITTER: James Collins

PROVIDER: E-GEOD-48282 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:STUDY THE EFFECT OF TGF beta ON SCHISTOSOME ADULT WORMSGoal: Primary: Assess the effects of TGF beta on transcription of adult male and female schistosomes. Secondary: Identify the pathways regulated by TGF beta in worm pairs, mature males, mature females, immature males and immature females.METHODS:Parasite: Schistosoma mansoni employed is the NMRI strain that is maintained in a Puerto Rican Biomphalaria glabrata and Golden Syrian hamsters. The life cycle is maintained in the LoVerde lab.Cercariae obtained from infected snails were used to infect experimental animals. The cercariae wre shed from snails exposed to multiple miracidia for mixed infections or from snails exposed to a single miracidia for single-sex infections. Schistosome worms were recovered from hamsters by perfusion (Duvall and DeWitt, 1967) 45 days post infection. Freshly perfused adult worms containing mixed-sex or single-sex infections were collected from infected golden hamsters, washed and incubated for overnight at 37C, 5% CO2 in MEM medium supplemented with 1 mM sodium pyruvate, 1 X non-essential amino acid solution, 1 X amino acid solution, 100 U ml penicillin, 100 ug ml streptomycin 1 ug ml amphotericin B (antibiotic antimycotic solution) and 2 mM Glutamax-I. Recombinant human TGF- beta1 was added individually to about 25 pairs or 30 individual schistosomes to a final concentrations of 1.0 nM and incubation was continued for another 24 hr. The controls were treated as above except no TGF-beta1 was added. The worms were placed in RNAlater and sent to WTSI (Anna) for RNA extraction and sequencing.Duvall RH, Dewitt WD. An improved perfusion technique for recovering adult schistosomes from laboratory animals. Am J Trop Med Hyg 1967;16:483-486.. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/

Project description:Schistosomiasis, caused by infection with Schistosoma trematode parasites, is one of the deadliest neglected tropical diseases in the world. The Schistosoma lifecycle involves the miracidia infection of an intermediate molluscan host, such as Biomphalaria glabrata. Dispersing B. glabrata attractant chemicals has been considered a method of minimising host infections. The attractiveness of B. glabrata is known to be reduced following infection; however, it remained unclear how the duration of infection affects attractiveness. Identifying excretory-secretory proteins (ESPs) abundant in attractive snail conditioned water (SCW) may facilitate the identification of attractants. This study obtained SCW ESPs from naïve snails and at different time-points post-miracidia exposure (PME; 16h-, 1 week, 2 weeks and 3 weeks), to identify those ESPs mediating Schistosome mansoni miracidia behavioural changes. The ESPs were quantitatively identified from all SCW samples using a label-free proteomic method and genome-derived protein databases of B. glabrata and S. mansoni. We found that changes in miracidia behaviour were similar upon exposure to naïve and 3W-PME SCW, including increased circling and quantity of miracidia in the field of view. Of the ESPs identified, 21 were shared in SCW of naïve and 3W-PME, and considered attractant candidates, including acetylcholine-binding protein and immune-related proteins such as biomphalysins and thioester-containing protein 1. Some biological processes were exclusive to attractant candidates, including obsolete pathogenesis and transmembrane transport activity. This suggests that compromised production of these proteins may diminish host attractiveness to miracidia. Additionally, several immune proteins were identified exclusively at 16-PME and 1W-PME, providing further information into changing B. glabrata immune responses throughout infection. These findings provide a list of attractant candidates to potentially decrease B. glabrata infection and minimise schistosomiasis.

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Functional genomic characterization of germinal cells in larval Schistosoma mansoni

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