Open Access Research article

Genome-wide expression profiling of in vivo-derived bloodstream parasite stages and dynamic analysis of mRNA alterations during synchronous differentiation in Trypanosoma brucei

Sarah Kabani1, Katelyn Fenn1, Alan Ross2, Al Ivens3, Terry K Smith4, Peter Ghazal2 and Keith Matthews1*

Author Affiliations

1 Centre for Immunity, Infection and Evolution, Institute of Immunology and Infection Research, School of Biological Sciences, King's Buildings, University of Edinburgh, Edinburgh, UK

2 Division of Pathway Medicine, University of Edinburgh Medical School, Chancellor's building, University of Edinburgh, Edinburgh, UK

3 Fios Genomics Ltd, ETTC, King's buildings, Edinburgh, UK

4 Centre for Biomolecular Sciences, The North Haugh, St Andrews University, UK

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BMC Genomics 2009, 10:427  doi:10.1186/1471-2164-10-427

Published: 11 September 2009

Abstract

Background

Trypanosomes undergo extensive developmental changes during their complex life cycle. Crucial among these is the transition between slender and stumpy bloodstream forms and, thereafter, the differentiation from stumpy to tsetse-midgut procyclic forms. These developmental events are highly regulated, temporally reproducible and accompanied by expression changes mediated almost exclusively at the post-transcriptional level.

Results

In this study we have examined, by whole-genome microarray analysis, the mRNA abundance of genes in slender and stumpy forms of T.brucei AnTat1.1 cells, and also during their synchronous differentiation to procyclic forms. In total, five biological replicates representing the differentiation of matched parasite populations derived from five individual mouse infections were assayed, with RNAs being derived at key biological time points during the time course of their synchronous differentiation to procyclic forms. Importantly, the biological context of these mRNA profiles was established by assaying the coincident cellular events in each population (surface antigen exchange, morphological restructuring, cell cycle re-entry), thereby linking the observed gene expression changes to the well-established framework of trypanosome differentiation.

Conclusion

Using stringent statistical analysis and validation of the derived profiles against experimentally-predicted gene expression and phenotypic changes, we have established the profile of regulated gene expression during these important life-cycle transitions. The highly synchronous nature of differentiation between stumpy and procyclic forms also means that these studies of mRNA profiles are directly relevant to the changes in mRNA abundance within individual cells during this well-characterised developmental transition.