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Patterns of gene expression in schistosomes: localization by whole mount in situ hybridization

  • G. P. DILLON (a1), J. C. ILLES (a2), H. V. ISAACS (a1) and R. A. WILSON (a1)

Summary

As a consequence of comprehensive transcriptome analysis followed by sequencing and draft assembly of the genome, the emphasis of schistosome research is shifting from the identification of genes to the characterization of their functions and interactions. Developmental biologists have long used whole mount in situ hybridization (WISH) to determine gene expression patterns, as a vital tool for formulating and testing hypotheses about function. This paper describes the application of WISH to the study of gene expression in larval and adult schistosomes. Fixed worms were permeablized by proteinase K treatment for hybridization with digoxygenin-labelled RNA probes, with binding being detected by alkaline phosphatase-coupled anti-digoxygenin antibodies, and BM Purple substrate. Discrete staining patterns for the transcripts of the molecules Sm29, cathepsin L, antigen 10.3 and chorion were observed in the tegument cell bodies, gut epithelium, oesophageal gland and vitelline lobules, respectively, of adult worms. Transcripts of the molecules SGTP4, GP18-22 and cathepsin L were localized to tegument cell bodies and embryonic gut, respectively, of lung schistosomula. We also showed that Fast Red TR fluorescent substrate can refine the pattern of localization permitting use of confocal microscopy. We believe that method of WISH will find broad application, in synergy with other emerging post-genomic techniques, such as RNA interference, to studies focused at increasing our molecular understanding of schistosomes.

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Corresponding author

*Corresponding author: Biology Department, University of York, Heslington, York YO10 5DD, UK. Tel: +44 (0) 1904 328592. Fax: +44 (0) 1904 328505. E-mail: gpd105@york.ac.uk

References

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Bogitsh, B. J., Dalton, J. P., Brady, C. P. and Brindley, P. J. (2001). Gut-associated immunolocalization of the Schistosoma mansoni cysteine proteases, SmCL1 and SmCL2. Journal of Parasitology 87, 237241.
Braschi, S., Borges, W. C. and Wilson, R. A. (2006). Proteomic analysis of the schistosome tegument and its surface membranes. Memórias do Instituto Oswaldo Cruz 101 (Suppl. 1), 205212.
Braschi, S., Curwen, R. S., Ashton, P. D., Verjovski-Almeida, S. and Wilson, R. A. (2005). The tegument surface membranes of the human blood parasite Schistosoma mansoni: a proteomic analysis after differential extraction. Proteomics 6, 14711482.
Braschi, S. and Wilson, R. A. (2006). Proteins exposed at the adult schistosome surface revealed by biotinylation. Molecular and Cellular Proteomics 5, 347356.
Cardoso, F. C., Pacifico, R. N., Mortara, R. A. and Oliveira, S. C. (2006). Human antibody responses of patients living in endemic areas for schistosomiasis to the tegumental protein Sm29 identified through genomic studies. Clinical and Experimental Immunology 144, 382391.
Coulson, P. S. (1997). The radiation-attenuated vaccine against schistosomes in animal models: paradigm for a human vaccine? Advances in Parasitology 39, 271336.
Cox, W. G. and Singer, V. L. (1999). A high-resolution, fluorescence-based method for localization of endogenous alkaline phosphatase activity. The Journal of Histochemistry and Cytochemistry 47, 14431456.
Curwen, R. S., Ashton, P. D., Sundaralingam, S. and Wilson, R. A. (2006). Identification of novel proteases and immunomodulators in the secretions of schistosome cercariae that facilitate host entry. Molecular and Cellular Proteomics 5, 835844.
Davis, R. E., Davis, A. H., Carroll, S. M., Rajkovic, A. and Rottman, F. M. (1988). Tandemly repeated exons encode 81-base repeats in multiple, developmentally regulated Schistosoma mansoni transcripts. Molecular and Biochemical Parasitology 8, 47454755.
Dillon, G. P., Feltwell, T., Skelton, J. P., Ashton, P. D., Coulson, P. S., Quail, M. A., Nikolaidou-Katsaridou, N., Wilson, R. A. and Ivens, A. C. (2006). Microarray analysis identifies genes preferentially expressed in the lung schistosomulum of Schistosoma mansoni. International Journal for Parasitology 36, 18.
el-Sherbeini, M., Ramadan, N., Bostian, K. A. and Knopf, P. M. (1991). Cloning and sequence analysis of the Schistosoma mansoni membrane glycoprotein antigen gene GP22. Molecular and Biochemical Parasitology 49, 8398.
Harland, R. M. (1991). In-situ hybridization: an improved whole-mount method for Xenopus embryos. Methods in Cell Biology 36, 685695.
Jones, K. J., Randall, L. M., McManus, D. P. and Engwerda, C. R. (2004). Laser microdissection microscopy in parasitology: microscopes meet thermocyclers. Trends in Parasitology 20, 502506.
Kelly, L. E., Nekkalapudi, S. and El Hodiri, H. M. (2007). Expression of the forkhead transcription factor FoxN4 in progenitor cells in the developing Xenopus laevis retina and brain. Gene Expression Patterns 7, 233238.
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, 209220.
Knobloch, J., Rossi, A., Osman, A., LoVerde, P. T., Klinkert, M. Q. and Grevelding, C. G. (2004). Cytological and biochemical evidence for a gonad-preferential interplay of SmFKBP12 and SmTbetaR-I in Schistosoma mansoni. Molecular and Biochemical Parasitology 138, 227236.
Knudsen, G. M., Medzihradszky, K. F., Lim, K. C., Hansell, E. and McKerrow, J. H. (2005). Proteomic analysis of Schistosoma mansoni cercarial secretions. Molecular and Cellular Proteomics 4, 18621875.
Koster, B., Dargatz, H., Schroder, J., Hirzmann, J., Haarmann, C., Symmons, P. and Kunz, W. (1988). Identification and localisation of the products of a putative eggshell precursor gene in the vitellarium of Schistosoma mansoni. Molecular and Biochemical Parasitology 31, 183198.
Lawrence, J. D. (1973). The ingestion of red blood cells by Schistosoma mansoni. The Journal of Parasitology 59, 6063.
Machado-Silver, J. R., Pelajo-Machado, M., Lenzi, H. L. and Gomes, D. C. (1998). Morphological study of adult male worms of Schistosoma mansoni Sambon, 1907 by confocal laser scanning microscopy. Memórias do Instituto Oswaldo Cruz 93 (Suppl. 1), 303307.
Michel, A., Ghoneim, H., Resto, M., Klinkert, M. Q. and Kunz, W. (1995). Sequence, characterization and localization of a cysteine proteinase cathepsin L in Schistosoma mansoni. Molecular and Biochemical Parasitology 73, 718.
Ming, Z., Dong, H., Zhong, Q., Grevelding, C. G. and Jiang, M. (2006). The effect of a mutagen (N-methyl-N-nitro-N-nitrosoguanidine) on cultured cells from adult Schistosoma japonicum. Parasitology Research 98, 430437.
Morris, G. P. and Threadgold, L. T. (1968). Ultrastructure of the tegument of adult Schistosoma mansoni. The Journal of Parasitology 54, 1527.
Osman, AHMED, Niles, G. and LoVerde, P. T. (2001). Identification and characterization of a Smad2 homologue from Schistosoma mansoni, a transforming growth factor-beta signal transducer. Journal of Biological Chemistry 276, 1007210082.
Pownall, M. E., Tucker, A. S., Slack, J. M. and Isaacs, H. V. (1996). eFGF, Xcad3 and Hox genes form a molecular pathway that establishes the anteroposterior axis in Xenopus. Development 122, 38813892.
Shaw, M. K. and Erasmus, D. A. (1982). Schistosoma mansoni: the presence and ultrastructure of vitelline cells in adult males. Journal of Helminthology 56, 5153.
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, 363369.
Skelly, P. J., Kim, J. W., Cunningham, J. and Shoemaker, C. B. (1994). Cloning, characterization, and functional expression of cDNAs encoding glucose transporter proteins from the human parasite Schistosoma mansoni. Journal of Biological Chemistry 269, 42474253.
Urieli-Shoval, S., Meek, R. L., Hanson, R. H., Ferguson, M., Gordon, D. and Benditt, E. P. (1992). Preservation of RNA for in situ hybridization: Carnoy's versus formaldehyde fixation. The Journal of Histochemistry and Cytochemistry 40, 18791885.
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, 148157.
Wilson, R. A., Ashton, P. D., Braschi, S., Dillon, G. P., Berriman, M. and Ivens, A. (2007). ‘Oming in on schistosomes: prospects and limitations for post-genomics’. Trends in Parasitology 23, 1420.
Wilson, R. A. and Barnes, P. E. (1974). The tegument of Schistosoma mansoni: observations on the formation, structure and composition of cytoplasmic inclusions in relation to tegument function. Parasitology 68, 239258.
Wilson, R. A. and Coulson, P. S. (2006). Schistosome vaccines: a critical appraisal. Memórias do Instituto Oswaldo Cruz 101 (Suppl. 1), 1320.

Keywords

Patterns of gene expression in schistosomes: localization by whole mount in situ hybridization

  • G. P. DILLON (a1), J. C. ILLES (a2), H. V. ISAACS (a1) and R. A. WILSON (a1)

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