Hostname: page-component-8448b6f56d-cfpbc Total loading time: 0 Render date: 2024-04-19T20:13:35.490Z Has data issue: false hasContentIssue false
  • English
  • Français

Culture for genetic manipulation of developmental stages of Schistosoma mansoni

Published online by Cambridge University Press:  21 September 2009

VICTORIA H. MANN ,
MARIA E. MORALES ,
GABRIEL RINALDI  and
PAUL J. BRINDLEY
VICTORIA H. MANN*
Affiliation:
Department of Microbiology, Immunology & Tropical Medicine, George Washington University Medical Center, Washington, DC20037USA
MARIA E. MORALES
Affiliation:
Tulane Cancer Center and Department of Epidemiology, Tulane University Health Sciences Center, New Orleans, Louisiana70112USA
GABRIEL RINALDI
Affiliation:
Department of Microbiology, Immunology & Tropical Medicine, George Washington University Medical Center, Washington, DC20037USA Departamento de Genética, Facultad de Medicina, Universidad de la República, (UDELAR), Montevideo, Uruguay.
PAUL J. BRINDLEY
Affiliation:
Department of Microbiology, Immunology & Tropical Medicine, George Washington University Medical Center, Washington, DC20037USA
*
*Corresponding author: Department of Microbiology, Immunology & Tropical Medicine, George Washington University Medical Center, Ross Hall Room 448, 2300 I Street, NW, Washington, DC20037USA. Fax +1 202 994 2913. E-mail: mtmvhm@gwumc.edu
Get access Rights & Permissions [Opens in a new window]

Summary

Genomes of the major human helminth parasites, and indeed many others of agricultural significance, are now the research focus of intensive genome sequencing and annotation. A draft genome sequence of the filarial parasite Brugia malayi was reported in 2007 and draft genomes of two of the human schistosomes, Schistosoma japonicum and S. mansoni reported in 2009. These genome data provide the basis for a comprehensive understanding of the molecular mechanisms involved in schistosome nutrition and metabolism, host-dependent development and maturation, immune evasion and invertebrate evolution. In addition, new potential vaccine candidates and drug targets will likely be predicted. However, testing these predictions is often not straightforward with schistosomes because of the difficulty and expense in maintenance of the developmental cycle. To facilitate this goal, several developmental stages can be maintained in vitro for shorter or longer intervals of time, and these are amenable to manipulation. Our research interests focus on experimental studies of schistosome gene functions, and more recently have focused on development of transgenesis and RNA interference with the longer term aim of heritable gene manipulation. Here we review methods to isolate and culture developmental stages of Schistosoma mansoni, including eggs, sporocysts, schistosomules and adults, in particular as these procedures relate to approaches for gene manipulation. We also discuss recent advances in genetic manipulation of schistosomes including the deployment of square wave electroporation to introduce reporter genes into cultured schistosomes.

Keywords

Schistosomesporocystschistosomuleeggin vitro cultureelectroporation
Type
Research Article
Information
Parasitology , Volume 137 , Special Issue 3: Development and applications of cestode and trematode laboratory models , March 2010 , pp. 451 - 462
Copyright
Copyright © Cambridge University Press 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Alfa, R. W. and Blesch, A. (2006). Murine and HIV-based retroviral vectors for in vitro and in vivo gene transfer. Methods in Molecular Medicine 129, 241–254. doi: 10.1385/1-59745-213-0:241.Google ScholarPubMed
Basch, P. F. (1981). Cultivation of Schistosoma mansoni in vitro. I. Establishment of cultures from cercariae and development until pairing. Journal of Parasitology 67, 179–185.CrossRefGoogle ScholarPubMed
Beckmann, S., Wippersteg, V., El-Bahay, A., Hirzmann, J., Oliveira, G. and Grevelding, C. G. (2007). Schistosoma mansoni: germ-line transformation approaches and actin-promoter analysis. Experimental Parasitology 117, 292–303. doi: 10.1016/j.exppara.2007.04.007.CrossRefGoogle ScholarPubMed
Berriman, M., Haas, B. J., LoVerde, P. T., Wilson, R. A., Dillon, G. P., Cerqueira, G. C., Mashiyama, S. T., Al-Lazikani, B., Andrade, L. F., Ashton, P. D., Aslett, M. A., Bartholomeu, D. C., Blandin, G., Caffrey, C. R., Coghlan, A., Coulson, R., Day, T. A., Delcher, A., DeMarco, R., Djikeng, A., Eyre, T., Gamble, J. A., Ghedin, E., Gu, Y., Hertz-Fowler, C., Hirai, H., Hirai, Y., Houston, R., Ivens, A., Johnston, D. A., Lacerda, D., Macedo, C. D., McVeigh, P., Ning, Z., Oliveira, G., Overington, J. P., Parkhill, J., Pertea, M., Pierce, R. J., Protasio, A. V., Quail, M. A., Rajandream, M. A., Rogers, J., Sajid, M., Salzberg, S. L., Stanke, M., Tivey, A. R., White, O., Williams, D. L., Wortman, J., Wu, W., Zamanian, M., Zerlotini, A., Fraser-Liggett, C. M., Barrell, B. G. and El-Sayed, N. M. (2009). The genome of the blood fluke Schistosoma mansoni. Nature 460, 352–358. doi: 10.1038/nature08160.CrossRefGoogle ScholarPubMed
Bixler, L. M., Lerner, J. P., Ivanchenko, M., McCormick, R. S., Barnes, D. W. and Bayne, C. J. (2001). Axenic culture of Schistosoma mansoni sporocysts in low O2 environments. Journal of Parasitology 87, 1167–1168.Google ScholarPubMed
Bouvard, V., Baan, R., Straif, K., Grosse, Y., Secretan, B., El Ghissassi, F., Benbrahim-Tallaa, L., Guha, N., Freeman, C., Galichet, L., Cogliano, V. and WHO International Agency for Research on Cancer Monograph Working Group (2009). A review of human carcinogens – Part B: biological agents. Lancet Oncology 10, 321–322.CrossRefGoogle ScholarPubMed
Brindley, P. J. and Pearce, E. J. (2007). Genetic manipulation of schistosomes. International Journal for Parasitology 37, 465–473. doi: 10.1016/j.ijpara.2006.12.012.CrossRefGoogle ScholarPubMed
Correnti, J. M., Brindley, P. J. and Pearce, E. J. (2005). Long-term suppression of cathepsin B levels by RNA interference retards schistosome growth. Molecular and Biochemical Parasitology 143, 209–215. doi: 10.1016/j.molbiopara.2005.06.007.CrossRefGoogle ScholarPubMed
Correnti, J. M., Jung, E., Freitas, T. C. and Pearce, E. J. (2007). Transfection of Schistosoma mansoni by electroporation and the description of a new promoter sequence for transgene expression. International Journal for Parasitology 37, 1107–1115. doi: 10.1016/j.ijpara.2007.02.011.CrossRefGoogle ScholarPubMed
Correnti, J. M. and Pearce, E. J. (2004). Transgene expression in Schistosoma mansoni: introduction of RNA into schistosomula by electroporation. Molecular and Biochemical Parasitology 137, 75–79. doi: 10.1016/j.molbiopara.2004.04.015.CrossRefGoogle ScholarPubMed
Coustau, C. and Yoshino, T. P. (2000). Flukes without snails: advances in the in vitro cultivation of intramolluscan stages of trematodes. Experimental Parasitology 94, 62–66. doi: 10.1006/expr.1999.4462.CrossRefGoogle ScholarPubMed
Crabb, B. S. and Gilson, P. R. (2007). A new system for rapid plasmid integration in Plasmodium parasites. Trends in Microbiology 15, 3–6. doi: 10.1016/j.tim.2006.11.006.CrossRefGoogle ScholarPubMed
Dalton, J. P., Day, S. R., Drew, A. C. and Brindley, P. J. (1997). A method for the isolation of schistosome eggs and miracidia free of contaminating host tissues. Parasitology 115, 29–32.CrossRefGoogle ScholarPubMed
Davis, H. E., Rosinski, M., Morgan, J. R. and Yarmush, M. L. (2004). Charged polymers modulate retrovirus transduction via membrane charge neutralization and virus aggregation. Biophysical Journal 86, 1234–1242. doi: 10.1016/S0006-3495(04)74197-1.CrossRefGoogle ScholarPubMed
Delcroix, M., Sajid, M., Caffrey, C. R., Lim, K. C., Dvorak, J., Hsieh, I., Bahgat, M., Dissous, C. and McKerrow, J. H. (2006). A multienzyme network functions in intestinal protein digestion by a platyhelminth parasite. Journal of Biological Chemistry 281, 39316–39329. doi: 10.1074/jbc.M607128200.CrossRefGoogle ScholarPubMed
Duvall, R. H. and DeWitt, W. B. (1967). An improved perfusion technique for recovering adult schistosomes from laboratory animals. American Journal of Tropical Medicine and Hygiene 16, 483–486.CrossRefGoogle ScholarPubMed
Freitas, T. C., Jung, E. and Pearce, E. J. (2007). TGF-beta signaling controls embryo development in the parasitic flatworm Schistosoma mansoni. PLoS Pathogens 3, e52. doi: 10.1371/journal.ppat.0030052.CrossRefGoogle ScholarPubMed
Gobert, G. N., Moertel, L., Brindley, P. J. and McManus, D. P. (2009). Developmental gene expression profiles of the human pathogen Schistosoma japonicum. BMC Genomics 10, 128. doi: 10.1186/1471-2164-10-128.CrossRefGoogle ScholarPubMed
Hackett, F. (1993). The culture of Schistosoma mansoni and production of life cycle stages. Methods in Molecular Biology (Clifton, N.J.) 21, 89–99. doi: 10.1385/0-89603-239-6:89.Google ScholarPubMed
Han, Z. G., Brindley, P. J., Wang, S. Y. and Chen, Z. (2009). Schistosome genetics: new perspectives on schistosome biology and host-parasite interaction. Annual Review of Genomics and Human Genetics doi: 10.1146/annurev-genom-082908-150036.CrossRefGoogle ScholarPubMed
Hewitson, J. P., Grainger, J. R. and Maizels, R. M. (2009). Helminth immunoregulation: the role of parasite secreted proteins in modulating host immunity. Molecular and Biochemical Parasitology 167, 1–11. doi: 10.1016/j.molbiopara.2009.04.008.CrossRefGoogle ScholarPubMed
Hirai, H., Taguchi, T., Saitoh, Y., Kawanaka, M., Sugiyama, H., Habe, S., Okamoto, M., Hirata, M., Shimada, M., Tiu, W. U., Lai, K., Upatham, E. S. and Agatsuma, T. (2000). Chromosomal differentiation of the Schistosoma japonicum complex. International Journal for Parasitology 30, 441–452.CrossRefGoogle ScholarPubMed
Hokke, C. H., Fitzpatrick, J. M. and Hoffmann, K. F. (2007). Integrating transcriptome, proteome and glycome analyses of Schistosoma biology. Trends in Parasitology 23, 165–174. doi: 10.1016/j.pt.2007.02.007.CrossRefGoogle ScholarPubMed
Hotez, P. J., Brindley, P. J., Bethony, J. M., King, C. H., Pearce, E. J. and Jacobson, J. (2008). Helminth infections: the great neglected tropical diseases. Journal of Clinical Investigation 118, 1311–1321. doi: 10.1172/JCI34261.CrossRefGoogle ScholarPubMed
Hu, W., Brindley, P. J., McManus, D. P., Feng, Z. and Han, Z. G. (2004). Schistosome transcriptomes: new insights into the parasite and schistosomiasis. Trends in Molecular Medicine 10, 217–225. doi: 10.1016/j.molmed.2004.03.002.CrossRefGoogle ScholarPubMed
Hu, W., Yan, Q., Shen, D. K., Liu, F., Zhu, Z. D., Song, H. D., Xu, X. R., Wang, Z. J., Rong, Y. P., Zeng, L. C., Wu, J., Zhang, X., Wang, J. J., Xu, X. N., Wang, S. Y., Fu, G., Zhang, X. L., Wang, Z. Q., Brindley, P. J., McManus, D. P., Xue, C. L., Feng, Z., Chen, Z. and Han, Z. G. (2003). Evolutionary and biomedical implications of a Schistosoma japonicum complementary DNA resource. Nature Genetics 35, 139–147. doi: 10.1038/ng1236.CrossRefGoogle ScholarPubMed
Ivanchenko, M. G., Lerner, J. P., McCormick, R. S., Toumadje, A., Allen, B., Fischer, K., Hedstrom, O., Helmrich, A., Barnes, D. W. and Bayne, C. J. (1999). Continuous in vitro propagation and differentiation of cultures of the intramolluscan stages of the human parasite Schistosoma mansoni. Proceedings of the National Academy of Sciences, USA 96, 4965–4970.CrossRefGoogle ScholarPubMed
James, E. R. and Taylor, M. G. (1976). Transformation of cercariae to schistosomula: a quantitative comparison of transformation techniques and of infectivity by different injection routes of the organisms produced. Journal of Helminthology 50, 223–233.CrossRefGoogle ScholarPubMed
Jolly, E. R., Chin, C. S., Miller, S., Bahgat, M. M., Lim, K. C., DeRisi, J. and McKerrow, J. H. (2007). Gene expression patterns during adaptation of a helminth parasite to different environmental niches. Genome Biology 8, R65. doi: 10.1186/gb-2007-8-4-r65.CrossRefGoogle ScholarPubMed
Kapp, K., Coustau, C., Wippersteg, V., Jourdane, J., Kunz, W. and Grevelding, C. G. (2003). Transplantation of in vitro-generated Schistosoma mansoni mother sporocysts into Biomphalaria glabrata. Parasitology Research 91, 482–485. doi: 10.1007/s00436-003-0951-1.Google ScholarPubMed
Kawanaka, M., Hayashi, S. and Ohtomo, H. (1983). A minimum essential medium for cultivation of Schistosoma japonicum eggs. Journal of Parasitology 69, 991–992.CrossRefGoogle ScholarPubMed
Kawanaka, M., Sidner, R. A. and Carter, C. E. (1985). In vitro transformation of Schistosoma japonicum miracidia to young sporocysts in a culture system for egg maturation. Journal of Parasitology 71, 368–370.CrossRefGoogle Scholar
Kines, K. J., Mann, V. H., Morales, M. E., Shelby, B. D., Kalinna, B. H., Gobert, G. N., Chirgwin, S. R. and Brindley, P. J. (2006). Transduction of Schistosoma mansoni by vesicular stomatitis virus glycoprotein-pseudotyped Moloney murine leukemia retrovirus. Experimental Parasitology 112, 209–220. doi: 10.1016/j.exppara.2006.02.003.CrossRefGoogle ScholarPubMed
Kines, K. J., Morales, M. E., Mann, V. H., Gobert, G. N. and Brindley, P. J. (2008). Integration of reporter transgenes into Schistosoma mansoni chromosomes mediated by pseudotyped murine leukemia virus. FASEB Journal 22, 2936–2948. doi: 10.1096/fj.08-108308.CrossRefGoogle ScholarPubMed
Lazdins, J. K., Stein, M. J., David, J. R. and Sher, A. (1982). Schistosoma mansoni: rapid isolation and purification of schistosomula of different developmental stages by centrifugation on discontinuous density gradients of Percoll. Experimental Parasitology 53, 39–44.CrossRefGoogle ScholarPubMed
Lewis, F. (Ed.) (1998). Schistosomiasis. Current Protocols in Immunology, Suppl. 28, In: Animal Models for Infectious Diseases. (ed. Coligan, J. E., Kruisbeek, A. M., Margulies, D. H., Shevach, E. M. and Strober, W.), Wiley, New York.Google Scholar
Liu, F., Chen, P., Cui, S. J., Wang, Z. Q. and Han, Z. G. (2008). SjTPdb: integrated transcriptome and proteome database and analysis platform for Schistosoma japonicum. BMC Genomics 9, 304. doi: 10.1186/1471-2164-9-304.CrossRefGoogle ScholarPubMed
Liu, F., Lu, J., Hu, W., Wang, S. Y., Cui, S. J., Chi, M., Yan, Q., Wang, X. R., Song, H. D., Xu, X. N., Wang, J. J., Zhang, X. L., Zhang, X., Wang, Z. Q., Xue, C. L., Brindley, P. J., McManus, D. P., Yang, P. Y., Feng, Z., Chen, Z. and Han, Z. G. (2006). New perspectives on host-parasite interplay by comparative transcriptomic and proteomic analyses of Schistosoma japonicum. PLoS Pathogens 2, e29. doi: 10.1371/journal.ppat.0020029.CrossRefGoogle ScholarPubMed
Mann, V. H., Morales, M. E., Kines, K. J. and Brindley, P. J. (2008). Transgenesis of schistosomes: approaches employing mobile genetic elements. Parasitology 135, 141–153. doi: 10.1017/S0031182007003824.CrossRefGoogle ScholarPubMed
Miskey, C., Izsvak, Z., Kawakami, K. and Ivics, Z. (2005). DNA transposons in vertebrate functional genomics. Cellular and Molecular Life Sciences 62, 629–641. doi: 10.1007/s00018-004-4232-7.CrossRefGoogle ScholarPubMed
Morales, M. E., Mann, V. H., Kines, K. J., Gobert, G. N., FraserM. J.,JR M. J.,JR, Kalinna, B. H., Correnti, J. M., Pearce, E. J. and Brindley, P. J. (2007). piggyBac transposon mediated transgenesis of the human blood fluke, Schistosoma mansoni. FASEB Journal 21, 3479–3489. doi: 10.1096/fj.07-8726com.CrossRefGoogle ScholarPubMed
Morales, M. E., Rinaldi, G., Gobert, G. N., Kines, K. J., Tort, J. F. and Brindley, P. J. (2008). RNA interference of Schistosoma mansoni cathepsin D, the apical enzyme of the hemoglobin proteolysis cascade. Molecular and Biochemical Parasitology 157, 160–168. doi: 10.1016/j.molbiopara.2007.10.009.CrossRefGoogle ScholarPubMed
Ndegwa, D., Krautz-Peterson, G. and Skelly, P. J. (2007). Protocols for gene silencing in schistosomes. Experimental Parasitology 117, 284–291. doi: 10.1016/j.exppara.2007.07.012.CrossRefGoogle ScholarPubMed
O'Brochta, D. A., Sethuraman, N., Wilson, R., Hice, R. H., Pinkerton, A. C., Levesque, C. S., Bideshi, D. K., Jasinskiene, N., Coates, C. J., James, A. A., Lehane, M. J. and Atkinson, P. W. (2003). Gene vector and transposable element behavior in mosquitoes. The Journal of Experimental Biology 206, 3823–3834.CrossRefGoogle ScholarPubMed
Pfeifer, A. and Hofmann, A. (2009). Lentiviral transgenesis. Methods in Molecular Biology (ed. Clifton, N. J.), 530, 391–405. doi: 10.1007/978-1-59745-471-1_21.Google Scholar
Plasterk, R. H., Izsvak, Z. and Ivics, Z. (1999). Resident aliens: the Tc1/mariner superfamily of transposable elements. Trends in Genetics 15, 326–332.CrossRefGoogle ScholarPubMed
Rinaldi, G., Morales, M. E., Alrefaei, Y. N., Cancela, M., Castillo, E., Dalton, J. P., Tort, J. F. and Brindley, P. J. (2009). RNA interference targeting leucine aminopeptidase blocks hatching of Schistosoma mansoni eggs. Molecular and Biochemical Parasitology 167, 118–126. doi: 10.1016/j.molbiopara.2009.05.002.CrossRefGoogle ScholarPubMed
Rinaldi, G., Morales, M. E., Cancela, M., Castillo, E., Brindley, P. J. and Tort, J. F. (2008). Development of functional genomic tools in trematodes: RNA interference and luciferase reporter gene activity in Fasciola hepatica. PLoS Neglected Tropical Diseases 2, e260. doi: 10.1371/journal.pntd.0000260.CrossRefGoogle ScholarPubMed
Sasaki, E., Suemizu, H., Shimada, A., Hanazawa, K., Oiwa, R., Kamioka, M., Tomioka, I., Sotomaru, Y., Hirakawa, R., Eto, T., Shiozawa, S., Maeda, T., Ito, M., Ito, R., Kito, C., Yagihashi,C. ,C., Kawai, K., Miyoshi, H., Tanioka, Y., Tamaoki, N., Habu, S., Okano, H. and Nomura, T. (2009). Generation of transgenic non-human primates with germline transmission. Nature 459, 523–527. doi: 10.1038/nature08090.CrossRefGoogle ScholarPubMed
Schistosoma japonicum Genome Sequencing and Functional Analysis Consortium, Liu, F., Zhou, Y., Wang, Z. Q., Lu, G., Zheng, H., Brindley, P. J., McManus, D. P., Blair, D., Zhang, Q. H., Zhong, Y., Wang, S., Han, Z. G. and Chen, Z. (2009). The Schistosoma japonicum genome reveals features of host-parasite interplay. Nature 460, 345–351. doi: 10.1038/nature08140.Google Scholar
Skelly, P. J., Da'dara, A. and Harn, D. A. (2003). Suppression of cathepsin B expression in Schistosoma mansoni by RNA interference. International Journal for Parasitology 33, 363–369.CrossRefGoogle ScholarPubMed
Smithers, S. R. and Terry, R. J. (1965). The infection of laboratory hosts with cercariae of Schistosoma mansoni and the recovery of the adult worms. Parasitology 55, 695–700.CrossRefGoogle ScholarPubMed
Stirewalt, M. and Lewis, F. A. (1981). Schistosoma mansoni: effect of rotifers on cercarial output, motility and infectivity. International Journal for Parasitology 11, 301–308.CrossRefGoogle ScholarPubMed
Terenius, O., Marinotti, O., Sieglaff, D. and James, A. A. (2008). Molecular genetic manipulation of vector mosquitoes. Cell Host and Microbe 4, 417–423. doi: 10.1016/j.chom.2008.09.002.CrossRefGoogle ScholarPubMed
Utzinger, J. and Keiser, J. (2004). Schistosomiasis and soil-transmitted helminthiasis: common drugs for treatment and control. Expert Opinion on Pharmacotherapy 5, 263–285. doi: 10.1517/14656566.5.2.263.CrossRefGoogle ScholarPubMed
Verjovski-Almeida, S., DeMarco, R., Martins, E. A., Guimaraes, P. E., Ojopi, E. P., Paquola, A. C., Piazza, J. P., Nishiyama, M. Y. Jr, Kitajima, J. P., Adamson, R. E., Ashton, P. D., Bonaldo, M. F., Coulson, P. S., Dillon, G. P., Farias, L. P., Gregorio, S. P., Ho, P. L., Leite, R. A., Malaquias, L. C., Marques, R. C., Miyasato, P. A., Nascimento, A. L., Ohlweiler, F. P., Reis, E. M., Ribeiro, M. A., Sa, R. G., Stukart, G. C., Soares, M. B., Gargioni, C., Kawano, T., Rodrigues, V., Madeira, A. M., Wilson, R. A., Menck, C. F., Setubal, J. C., Leite, L. C. and Dias-Neto, E. (2003). Transcriptome analysis of the acoelomate human parasite Schistosoma mansoni. Nature Genetics 35, 148–157. doi: 10.1038/ng1237.CrossRefGoogle ScholarPubMed