The uptake of Texas Red-BSA in the excretory system of schistosomes and its colocalisation with ER60 promoter-induced GFP in transiently transformed adult males.
The excretory system of schistosomes has focused some attention during the last years since accumulating evidence suggests that it plays an important role in the host-parasite interaction. Signalling molecules such as phosphatases, but also proteases have been localised in the excretory system. To some extent, however, localisation studies are limited by the fact that sections of fixed specimens are used. In this study, we tested the fluorescent molecules FITC-dextran and Texas Red-BSA for their ability to enter the excretory system of living Schistosoma mansoni males. It is demonstrated that the dyes selectively stain the excretory tubules which are widely distributed along the worm body. This finding was used to investigate whether the staining of worms with Texas Red-BSA can help to localise transgene activity in worms which were transiently transformed by particle bombardment. A vector was used for transformation which contained the green fluorescent protein gene, under the control of the regulatory elements of the cysteine protease ER60 gene. After transformation and staining, confocal laser scanning microscopy revealed that ER60-induced green fluorescent protein activity colocalises with Texas Red-BSA in the excretory tubules. The results suggest a role for ER60 during the host-parasite interaction. Furthermore, the colocalisation approach introduced here opens further perspectives to characterise gene-expression profiles in this parasite.
D000818:Animals; D019470:Biolistics / Q000379:methods; D003546:Cysteine Endopeptidases / Q000235:genetics; D003911:Dextrans / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives* / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D005822:Genetic Vectors / Q000008:administration & dosage; D049452:Green Fluorescent Proteins; D007413:Intestinal Mucosa / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D011401:Promoter Regions, Genetic; D012550:Schistosoma mansoni / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic; D014966:Xanthenes / Q000378:metabolism*
D003911:Dextrans; D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D014966:Xanthenes; C015219:fluorescein isothiocyanate dextran; D049452:Green Fluorescent Proteins; C034657:Texas red; D003546:Cysteine Endopeptidases; D016650:Fluorescein-5-isothiocyanate