Skip to main content

Methods for Studying the Germline of the Human Parasite Schistosoma mansoni

  • Protocol
  • First Online:
Germline Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1463))

Abstract

Schistosomes are flatworm parasites that claim the lives of more than 200,000 people in poverty-stricken regions every year. Much of the pathology due to infection is the direct result of injury spurred by the parasite’s eggs becoming lodged in host tissues. Thus, asking basic questions about germ cell biology may not only identify novel therapeutic approaches, but could also uncover conserved mechanisms that regulate the germline in diverse metazoa. Here, we detail useful methods for studying the schistosome germline including EdU labeling, whole-mount in situ hybridization, and RNA interference. These methods will hopefully lead to new insights about germline development in the schistosome and facilitate new investigators to begin asking questions about these important and fascinating parasites.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Chitsulo L, Engels D, Montresor A, Savioli L (2000) The global status of schistosomiasis and its control. Acta Trop 77(1):41–51, doi:S0001-706X(00)00122-4 [pii]

    Article  CAS  PubMed  Google Scholar 

  2. van der Werf MJ, de Vlas SJ, Brooker S, Looman CW, Nagelkerke NJ, Habbema JD, Engels D (2003) Quantification of clinical morbidity associated with schistosome infection in sub-Saharan Africa. Acta Trop 86(2-3):125–139

    Article  PubMed  Google Scholar 

  3. Basch PF (1991) Schistosomes: development, reproduction, and host relations. Oxford University Press, New York

    Google Scholar 

  4. Pearce EJ, MacDonald AS (2002) The immunobiology of schistosomiasis. Nat Rev Immunol 2(7):499–511. doi:10.1038/nri843

    Article  CAS  PubMed  Google Scholar 

  5. Skinner DE, Rinaldi G, Koziol U, Brehm K, Brindley PJ (2014) How might flukes and tapeworms maintain genome integrity without a canonical piRNA pathway? Trends Parasitol 30(3):123–129. doi:10.1016/j.pt.2014.01.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Skinner DE, Rinaldi G, Suttiprapa S, Mann VH, Smircich P, Cogswell AA, Williams DL, Brindley PJ (2012) Vasa-like DEAD-Box RNA helicases of Schistosoma mansoni. PLoS Negl Trop Dis 6(6):e1686. doi:10.1371/journal.pntd.0001686, PNTD-D-12-00056 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Tsai IJ, Zarowiecki M, Holroyd N, Garciarrubio A, Sanchez-Flores A, Brooks KL, Tracey A, Bobes RJ, Fragoso G, Sciutto E, Aslett M, Beasley H, Bennett HM, Cai J, Camicia F, Clark R, Cucher M, De Silva N, Day TA, Deplazes P, Estrada K, Fernandez C, Holland PW, Hou J, Hu S, Huckvale T, Hung SS, Kamenetzky L, Keane JA, Kiss F, Koziol U, Lambert O, Liu K, Luo X, Luo Y, Macchiaroli N, Nichol S, Paps J, Parkinson J, Pouchkina-Stantcheva N, Riddiford N, Rosenzvit M, Salinas G, Wasmuth JD, Zamanian M, Zheng Y, Taenia solium Genome C, Cai X, Soberon X, Olson PD, Laclette JP, Brehm K, Berriman M (2013) The genomes of four tapeworm species reveal adaptations to parasitism. Nature 496 (7443):57–63. Doi: 10.1038/nature12031

    Google Scholar 

  8. Wang Y, Stary JM, Wilhelm JE, Newmark PA (2010) A functional genomic screen in planarians identifies novel regulators of germ cell development. Genes Dev 24(18):2081–2092. doi:10.1101/gad.1951010, 24/18/2081 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Newmark PA, Wang Y, Chong T (2008) Germ cell specification and regeneration in planarians. Cold Spring Harb Symp Quant Biol 73:573–581. doi:10.1101/sqb.2008.73.022, sqb.2008.73.022 [pii]

    Article  CAS  PubMed  Google Scholar 

  10. Collins JJ III, King RS, Cogswell A, Williams DL, Newmark PA (2011) An atlas for Schistosoma mansoni organs and life-cycle stages using cell type-specific markers and confocal microscopy. PLoS Negl Trop Dis 5(3):e1009. doi:10.1371/journal.pntd.0001009

    Article  PubMed  PubMed Central  Google Scholar 

  11. Collins JJ III, Wang B, Lambrus BG, Tharp ME, Iyer H, Newmark PA (2013) Adult somatic stem cells in the human parasite Schistosoma mansoni. Nature 494(7438):476–479. doi:10.1038/nature11924

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Beckmann S, Buro C, Dissous C, Hirzmann J, Grevelding CG (2010) The Syk kinase SmTK4 of Schistosoma mansoni is involved in the regulation of spermatogenesis and oogenesis. PLoS Pathog 6(2):e1000769. doi:10.1371/journal.ppat.1000769

    Article  PubMed  PubMed Central  Google Scholar 

  13. Quack T, Knobloch J, Beckmann S, Vicogne J, Dissous C, Grevelding CG (2009) The formin-homology protein SmDia interacts with the Src kinase SmTK and the GTPase SmRho1 in the gonads of Schistosoma mansoni. PLoS One 4(9):e6998. doi:10.1371/journal.pone.0006998

    Article  PubMed  PubMed Central  Google Scholar 

  14. Hahnel 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. doi:10.1371/journal.pntd.0002336

    Article  PubMed  PubMed Central  Google Scholar 

  15. Cogswell AA, Collins JJ III, Newmark PA, Williams DL (2011) Whole mount in situ hybridization methodology for Schistosoma mansoni. Mol Biochem Parasitol 178(1-2):46–50. doi:10.1016/j.molbiopara.2011.03.001, S0166-6851(11)00091-0 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Pearson BJ, Eisenhoffer GT, Gurley KA, Rink JC, Miller DE, Sánchez Alvarado A (2009) Formaldehyde-based whole-mount in situ hybridization method for planarians. Dev Dyn 238(2):443–450. doi:10.1002/dvdy.21849

    Article  PubMed  PubMed Central  Google Scholar 

  17. King RS, Newmark PA (2013) In situ hybridization protocol for enhanced detection of gene expression in the planarian Schmidtea mediterranea. BMC Dev Biol 13:8. doi:10.1186/1471-213X-13-8

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Stefanic S, Dvorak J, Horn M, Braschi S, Sojka D, Ruelas DS, Suzuki B, Lim KC, Hopkins SD, McKerrow JH, Caffrey CR (2010) RNA Interference in Schistosoma mansoni schistosomula: selectivity, sensitivity and operation for larger-scale screening. PLoS Negl Trop Dis 4(10):e850. doi:10.1371/journal.pntd.0000850

    Article  PubMed  PubMed Central  Google Scholar 

  19. Skelly PJ, Da'dara A, Harn DA (2003) Suppression of cathepsin B expression in Schistosoma mansoni by RNA interference. Int J Parasitol 33(4):363–369, doi:S0020751903000304 [pii]

    Article  CAS  PubMed  Google Scholar 

  20. Boyle JP, Wu XJ, Shoemaker CB, Yoshino TP (2003) Using RNA interference to manipulate endogenous gene expression in Schistosoma mansoni sporocysts. Mol Biochem Parasitol 128(2):205–215, doi: S0166685103000781 [pii]

    Article  CAS  PubMed  Google Scholar 

  21. Rouhana L, Weiss JA, Forsthoefel DJ, Lee H, King RS, Inoue T, Shibata N, Agata K, Newmark PA (2013) RNA interference by feeding in vitro-synthesized double-stranded RNA to planarians: methodology and dynamics. Dev Dyn 242(6):718–730. doi:10.1002/dvdy.23950

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Lewis FA, Liang YS, Raghavan N, Knight M (2008) The NIH-NIAID schistosomiasis resource center. PLoS Negl Trop Dis 2(7):e267. doi:10.1371/journal.pntd.0000267

    Article  PubMed  PubMed Central  Google Scholar 

  23. Collins JJ III, Hou X, Romanova EV, Lambrus BG, Miller CM, Saberi A, Sweedler JV, Newmark PA (2010) Genome-wide analyses reveal a role for peptide hormones in planarian germline development. PLoS Biol 8(10)

    Google Scholar 

  24. He B, Rong M, Lyakhov D, Gartenstein H, Diaz G, Castagna R, McAllister WT, Durbin RK (1997) Rapid mutagenesis and purification of phage RNA polymerases. Protein Expr Purif 9(1):142–151. doi:10.1006/prep.1996.0663, S1046-5928(96)90663-4 [pii]

    Article  CAS  PubMed  Google Scholar 

  25. Sive HL, Grainger RM, Harland RM (2007) Baskets for in situ hybridization and immunohistochemistry. CSH Protoc 2007: pdb prot4777. doi: 10.1101/pdb.prot4777

    Google Scholar 

  26. Basch PF (1981) Cultivation of Schistosoma mansoni in vitro. I Establishment of cultures from cercariae and development until pairing. J Parasitol 67(2):179–185

    Article  CAS  PubMed  Google Scholar 

  27. Collins JJ III, Newmark PA (2013) It's no fluke: the planarian as a model for understanding schistosomes. PLoS Pathog 9(7):e1003396. doi:10.1371/journal.ppat.1003396

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to James J. Collins III .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media New York

About this protocol

Cite this protocol

Collins, J.N.R., Collins, J.J. (2017). Methods for Studying the Germline of the Human Parasite Schistosoma mansoni . In: Buszczak, M. (eds) Germline Stem Cells. Methods in Molecular Biology, vol 1463. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4017-2_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-4017-2_2

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-4015-8

  • Online ISBN: 978-1-4939-4017-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics