RNA-Seq analysis validates the use of culture-derived Trypanosoma brucei and provides new markers for mammalian and insect life-cycle stages.

Naguleswaran, Arunasalam; Doiron, Nicholas; Roditi, Isabel (2018). RNA-Seq analysis validates the use of culture-derived Trypanosoma brucei and provides new markers for mammalian and insect life-cycle stages. BMC Genomics, 19(1), p. 227. BioMed Central 10.1186/s12864-018-4600-6

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BACKGROUND Trypanosoma brucei brucei, the parasite causing Nagana in domestic animals, is closely related to the parasites causing sleeping sickness, but does not infect humans. In addition to its importance as a pathogen, the relative ease of genetic manipulation and an innate capacity for RNAi extend its use as a model organism in cell and infection biology. During its development in its mammalian and insect (tsetse fly) hosts, T. b. brucei passes through several different life-cycle stages. There are currently four life-cycle stages that can be cultured: slender forms and stumpy forms, which are equivalent to forms found in the mammal, and early and late procyclic forms, which are equivalent to forms in the tsetse midgut. Early procyclic forms show coordinated group movement (social motility) on semi-solid surfaces, whereas late procyclic forms do not. RESULTS RNA-Seq was performed on biological replicates of each life-cycle stage. These constitute the first datasets for culture-derived slender and stumpy bloodstream forms and early and late procyclic forms. Expression profiles confirmed that genes known to be stage-regulated in the animal and insect hosts were also regulated in culture. Sequence reads of 100-125 bases provided sufficient precision to uncover differential expression of closely related genes. More than 100 transcripts showed peak expression in stumpy forms, including adenylate cyclases and several components of inositol metabolism. Early and late procyclic forms showed differential expression of 73 transcripts, a number of which encoded proteins that were previously shown to be stage-regulated. Moreover, two adenylate cyclases previously shown to reduce social motility are up-regulated in late procyclic forms. CONCLUSIONS This study validates the use of cultured bloodstream forms as alternatives to animal-derived parasites and yields new markers for all four stages. In addition to underpinning recent findings that early and late procyclic forms are distinct life-cycle stages, it could provide insights into the reasons for their different biological properties.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Department of Biology > Institute of Cell Biology

UniBE Contributor:

Naguleswaran, Arunasalam; Doiron, Nicholas and Roditi, Isabel

Subjects:

500 Science > 570 Life sciences; biology

ISSN:

1471-2164

Publisher:

BioMed Central

Funders:

[UNSPECIFIED] Howard Hughes Medical institute
[4] Swiss National Science Foundation
[UNSPECIFIED] Canton of Bern

Language:

English

Submitter:

Isabel Roditi

Date Deposited:

19 Jun 2018 15:08

Last Modified:

24 Jun 2018 02:26

Publisher DOI:

10.1186/s12864-018-4600-6

PubMed ID:

29606092

Uncontrolled Keywords:

3R Amino acid transporters Cation transporters Culture-derived RNA-Seq Replacement of animal models Stage-specific Transcriptome Trypanosome Tsetse

BORIS DOI:

10.7892/boris.117300

URI:

https://boris.unibe.ch/id/eprint/117300

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