Dillon, G.P., Illesh, J.C., Isaacs, H.V. and Wilson, R.A. (2007) Patterns of gene expression in schistosomes: localization by whole mount in situ hybridization. Parasitology. pp. 1589-1597. ISSN 0031-1820Full text available as:
rom 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.
|Copyright, Publisher and Additional Information:||Copyright © Cambridge University Press 2007. Reproduced in accordance with the publisher's self archiving policy. Available from August 2008.|
|Keywords:||Schistosoma mansoni, RNA probe, confocal microscopy|
|Academic Units:||The University of York > Biology (York)|
|Depositing User:||Repository Officer|
|Date Deposited:||24 Jan 2008 16:25|
|Last Modified:||17 Oct 2013 14:13|