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14-3-3- and II/3-10-gene expression as molecular markers to address viability and growth activity of Echinococcus multilocularis metacestodes

Published online by Cambridge University Press:  12 September 2005

J. MATSUMOTO ,
N. MÜLLER ,
A. HEMPHILL ,
Y. OKU ,
M. KAMIYA  and
B. GOTTSTEIN
J. MATSUMOTO
Affiliation:
Institute of Parasitology, University of Bern, Länggass-Strasse 122, CH-3001 Bern, Switzerland Laboratory of Parasitology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan
N. MÜLLER
Affiliation:
Institute of Parasitology, University of Bern, Länggass-Strasse 122, CH-3001 Bern, Switzerland
A. HEMPHILL
Affiliation:
Institute of Parasitology, University of Bern, Länggass-Strasse 122, CH-3001 Bern, Switzerland
Y. OKU
Affiliation:
Laboratory of Parasitology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan
M. KAMIYA
Affiliation:
Laboratory of Parasitology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan Present address: Laboratory of Environmental Zoology, Department of Biosphere and Environmental Sciences, Faculty of Environment Systems, Rakuno Gakuen University, Bunkyodai-Midorimachi, Ebetsu 069-8501, Japan.
B. GOTTSTEIN
Affiliation:
Institute of Parasitology, University of Bern, Länggass-Strasse 122, CH-3001 Bern, Switzerland
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Abstract

The present study aimed to search for and characterize parasite molecules, whose expression levels correlate with the viability and growth activity of Echinococcus multilocularis metacestodes. We focused on the expression profiles of 2 parasite-derived genes, 14-3-3 and II/3-10, as putative molecular markers for viability and growth activity of the larval parasite. In experiments in vivo, gene expression levels of 14-3-3 and II/3-10 were relatively quantified by real-time reverse transcription-PCR using a housekeeping gene, beta-actin, as a reference reaction. All three reactions were compared with growth activity of the parasite developing in permissive nu/nu and in non-permissive wild type BALB/c mice. At 2 months p.i., the transcription level of 14-3-3 was significantly higher in parasites actively proliferating in nu/nu mice compared to parasites moderately growing in wild type mice. Immunoblotting experiments confirmed at the protein level that 14-3-3 was over-expressed in parasites derived from nu/nu mice at 2 months p.i. In vitro treatment of E. multilocularis with an anti-echinococcal drug nitazoxanide resulted in a significant decrease of both 14-3-3 and II/3-10 transcription levels found after 8 days of treatment, which correlated with the kinetics of a housekeeping gene, beta-actin. The conclusion is that 14-3-3, combined with II/3-10, exhibits good potential as a molecular marker to assess viability and growth activity of the parasite.

Keywords

Echinococcus multilocularisviabilitygrowth activity14-3-3II/3-10
Type
Research Article
Information
Parasitology , Volume 132 , Issue 1 , January 2006 , pp. 83 - 94
Copyright
© 2005 Cambridge University Press

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