Quantitative assessment of DNA replication to monitor microgametogenesis in Plasmodium berghei

Targeting the crucial step of Plasmodium transition from vertebrate host to mosquito vector is a promising approach to eliminate malaria. Uptake by the mosquito activates gametocytes within seconds, and in the case of male (micro) gametocytes leads to rapid DNA replication and the release of eight f...

Full description

Saved in:
Bibliographic Details
Published inMolecular and biochemical parasitology Vol. 168; no. 2; pp. 172 - 176
Main Authors Raabe, Andreas C., Billker, Oliver, Vial, Henri J., Wengelnik, Kai
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 01.12.2009
Amsterdam: Elsevier
Elsevier
Elsevier/North-Holland Biomedical Press
Subjects
Animals
DNA
DNA Replication
DNA, Protozoan - biosynthesis
Exflagellation
Female
Gametocyte
Hypoxanthine - metabolism
Life Sciences
Malaria
Male
Microbiology and Parasitology
Parasitology
Plasmodium berghei
Plasmodium berghei - genetics
Plasmodium berghei - growth & development
Plasmodium berghei/genetics/growth & development
Protozoan/biosynthesis
Staining and Labeling - methods
Transmission
Tritium - metabolism
Gametocyte
Malaria
Exflagellation
Transmission
Online AccessGet full text

Cover

More Information
Summary:Targeting the crucial step of Plasmodium transition from vertebrate host to mosquito vector is a promising approach to eliminate malaria. Uptake by the mosquito activates gametocytes within seconds, and in the case of male (micro) gametocytes leads to rapid DNA replication and the release of eight flagellated gametes. We developed a sensitive assay to monitor P. berghei microgametocyte activation based on [3H]hypoxanthine incorporation into DNA. Optimal pH range and xanthurenic acid concentrations for gametocyte activation were established and the kinetics of DNA replication investigated. Significance of the method was confirmed using P. berghei mutants and the assay was applied to analyse the effect of protease inhibitors, which revealed differences regarding their inhibitory action. The developed method thus appears suitable for reproducible determination of microgametocyte activation, medium-throughput drug screenings and deeper investigation of early blocks in gametogenesis and will facilitate the analysis of compounds for transmission blocking activities.
Bibliography:http://dx.doi.org/10.1016/j.molbiopara.2009.08.004
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
PMCID: PMC2789244
ISSN:0166-6851
1872-9428
1872-9428
DOI:10.1016/j.molbiopara.2009.08.004