Salivary gland-specific P. berghei reporter lines enable rapid evaluation of tissue-specific sporozoite loads in mosquitoes

Ramakrishnan, C., Rademacher, A., Soichot, J., Costa, G., Waters, A.P. , Janse, C.J., Ramesar, J., Franke-Fayard, B.M. and Levashina, E.A. (2012) Salivary gland-specific P. berghei reporter lines enable rapid evaluation of tissue-specific sporozoite loads in mosquitoes. PLoS ONE, 7(5), e36376. (doi: 10.1371/journal.pone.0036376)

Abstract

Malaria is a life-threatening human infectious disease transmitted by mosquitoes. Levels of the salivary gland sporozoites (sgs), the only mosquito stage infectious to a mammalian host, represent an important cumulative index of <i>Plasmodium</i> development within a mosquito. However, current techniques of sgs quantification are laborious and imprecise. Here, transgenic <i>P. berghei</i> reporter lines that produce the green fluorescent protein fused to luciferase (GFP-LUC) specifically in sgs were generated, verified and characterised. Fluorescence microscopy confirmed the sgs stage specificity of expression of the reporter gene. The luciferase activity of the reporter lines was then exploited to establish a simple and fast biochemical assay to evaluate sgs loads in whole mosquitoes. Using this assay we successfully identified differences in sgs loads in mosquitoes silenced for genes that display opposing effects on <i>P. berghei</i> ookinete/oocyst development. It offers a new powerful tool to study infectivity of <i>P. berghei</i> to the mosquito, including analysis of vector-parasite interactions and evaluation of transmission-blocking vaccines.

• Item Type: Articles
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Waters, Professor Andy
• Authors: Ramakrishnan, C., Rademacher, A., Soichot, J., Costa, G., Waters, A.P., Janse, C.J., Ramesar, J., Franke-Fayard, B.M., and Levashina, E.A.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: PLoS ONE
• Publisher: Public Library of Science
• ISSN: 1932-6203
• Published Online: 04 May 2012
• Copyright Holders: Copyright © 2012 the authors
• First Published: First published in PLoS One 7 (5): e36376
• Publisher Policy: Reproduced in accordance with the copyright policy of the publisher