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Promiscuous Expression of alpha-Tubulin II in Maturing Male and Female Plasmodium falciparum Gametocytes

Schwank, S; Sutherland, CJ; Drakeley, CJ; (2010) Promiscuous Expression of alpha-Tubulin II in Maturing Male and Female Plasmodium falciparum Gametocytes. PLOS ONE , 5 (12) , Article e14470. 10.1371/journal.pone.0014470. Green open access

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Abstract

Background: Antimalarial interventions designed to impact on the transmissible sexual stages of Plasmodium falciparum are evaluated by measurement of peripheral gametocyte carriage in vivo and infectivity to mosquitoes. Drug or vaccine-elicited effects may differentially affect the relative abundance of mature male and female sexual forms, and this can be measured by estimation of sex ratios before and after intervention in vivo and in vitro. Measuring the impact of anti-gametocyte drugs on sexual commitment of immature gametocyte stages in vitro is not currently possible as male and female parasites cannot be distinguished by morphology alone prior to stage IV.Methodology/Principal Findings: We have modified an existing immunofluorescence-based approach for distinguishing male and female gametocytes during development in vitro, by using highly synchronised magnetically-enriched gametocyte preparations at different stages of maturity. Antibodies recognising alpha-tubulin II ( males) and Pfg377 ( females) were used to attempt to discriminate the sexes. Transcription of these two proteins was not coordinated during in vitro development, with pfg377 transcripts accumulating only late in development, immediately prior to immunofluorescent signals from the PfG377 protein appearing in stage IV gametocytes. Contrary to previous descriptions of this protein as male-specific in P. falciparum, alpha-tubulin II recognised both male and female gametocytes at stages I to IV, but evidence of differential expression levels of this protein in late stage male and female gametocytes was found. Using antibodies recognising PfG377 as the primary marker and alpha-tubulin II as a secondary marker, robust estimates of sex ratio in in vitro cultures were obtained for gametocytes at stage IV or later, and validated by light microscopic counts. However, sex ratio estimation was not possible for early stage gametocytes due to the promiscuity of alpha-tubulin II protein expression, and the relatively late accumulation of PfG377 during the development process.Conclusions/Significance: This approach is a feasible method for the evaluation of drug impacts on late-stage gametocyte sex ratio in in vitro studies. Additional sex-specific antigens need to be evaluated for sex ratio estimation in early stage gametocyte preparations.

Type: Article
Title: Promiscuous Expression of alpha-Tubulin II in Maturing Male and Female Plasmodium falciparum Gametocytes
Open access status: An open access version is available from UCL Discovery
DOI: 10.1371/journal.pone.0014470
Publisher version: http://dx.doi.org/10.1371/journal.pone.0014470
Language: English
Additional information: © 2010 Schwank et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The work was supported by a PhD Studentship to SS, awarded by the Europaeischer Landersverein der Industrie. CJS was supported by the UK Health Protection Agency. CJD was supported by Wellcome Trust grant #063516. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Keywords: HUMAN MALARIA PARASITE, SEX-RATIO, OSMIOPHILIC BODIES, SEQUENCE-ANALYSIS, BLOOD STAGES, COMMITMENT, PROTEIN, PFG377, GENES, STAGE
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Infection and Immunity
URI: https://discovery.ucl.ac.uk/id/eprint/1309945
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