Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single-cell transcriptomics

African trypanosomes proliferate as bloodstream forms (BSFs) and procyclic forms in the mammal and tsetse fly midgut, respectively. This allows them to colonise the host environment upon infection and ensure life cycle progression. Yet, understanding of the mechanisms that regulate and drive the cel...

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Bibliographic Details
Published in:eLife 2023-05, Vol.12
Main Authors: Briggs, Emma M, Marques, Catarina A, Oldrieve, Guy R, Hu, Jihua, Otto, Thomas D, Matthews, Keith R
Format: Article
Language:English
Subjects:
Cell Biology
Cell cycle
Comparative analysis
Cryopreservation
Exocrine glands
Freeze-thawing
Genes
Genomes
Genomics
Glycerol
Health aspects
Kinases
Life cycles
Microbiology and Infectious Disease
Midgut
Motility
Parasites
Phase transitions
Protozoa
Protozoan Proteins - analysis
RNA, Protozoan - analysis
Single-Cell Gene Expression Analysis
single-cell transcriptomics
Transcriptomes
Transcriptomics
Trypanosoma - cytology
Trypanosoma - growth & development
Trypanosoma - metabolism
Trypanosoma brucei
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Summary:African trypanosomes proliferate as bloodstream forms (BSFs) and procyclic forms in the mammal and tsetse fly midgut, respectively. This allows them to colonise the host environment upon infection and ensure life cycle progression. Yet, understanding of the mechanisms that regulate and drive the cell replication cycle of these forms is limited. Using single-cell transcriptomics on unsynchronised cell populations, we have obtained high resolution cell cycle regulated (CCR) transcriptomes of both procyclic and slender BSF without prior cell sorting or synchronisation. Additionally, we describe an efficient freeze-thawing protocol that allows single-cell transcriptomic analysis of cryopreserved . Computational reconstruction of the cell cycle using periodic pseudotime inference allowed the dynamic expression patterns of cycling genes to be profiled for both life cycle forms. Comparative analyses identify a core cycling transcriptome highly conserved between forms, as well as several genes where transcript levels dynamics are form specific. Comparing transcript expression patterns with protein abundance revealed that the majority of genes with periodic cycling transcript and protein levels exhibit a relative delay between peak transcript and protein expression. This work reveals novel detail of the CCR transcriptomes of both forms, which are available for further interrogation via an interactive webtool.
ISSN:2050-084X
2050-084X
DOI:10.7554/eLife.86325