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Here you can explore data from tsetse stages of T. brucei. Clustering analysis was performed in SC3. For more details on the data and analysis see our paper.
Virginia M. Howick, Lori Peacock, Chris Kay, Clare Collett, Wendy Gibson, Mara K.N. Lawniczak
Abstract
Early diverging lineages such as trypanosomes can provide clues to the evolution of sexual
reproduction in eukaryotes. In Trypanosoma brucei, the pathogen that causes Human
African Trypanosomiasis, sexual reproduction occurs during the part of the parasite’s life
cycle in the salivary glands of the insect host, but analysis of the molecular signatures that
define these sexual forms is complicated because they mingle with more numerous,
mitotically-dividing developmental stages. We used single-cell RNA-sequencing (scRNAseq)
to profile 388 individual trypanosomes from midgut, proventriculus, and salivary glands of
infected tsetse flies allowing us to identify tissue-specific cell types. Further investigation of
salivary gland parasite transcriptomes revealed fine-scale changes in gene expression over a
developmental progression from putative sexual forms through metacyclics expressing
variant surface glycoprotein genes via attached epimastigotes. The cluster of cells
potentially containing sexual forms was characterized by high expression of the gamete
fusion protein HAP2, together with an array of surface proteins and several genes of
unknown function. We linked these expression patterns to distinct morphological forms
using immunofluorescence assays and reporter gene expression to demonstrate that the
kinetoplastid-conserved gene Tb927.10.12080 is exclusively expressed at high levels by
meiotic intermediates and gametes. We speculate that this protein, currently of unknown
function, plays a role in gamete formation and/or fusion.