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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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Autores principales: Briggs, Emma M, Marques, Catarina A, Oldrieve, Guy R, Hu, Jihua, Otto, Thomas D, Matthews, Keith R
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10212563/
https://www.ncbi.nlm.nih.gov/pubmed/37166108
http://dx.doi.org/10.7554/eLife.86325
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author Briggs, Emma M
Marques, Catarina A
Oldrieve, Guy R
Hu, Jihua
Otto, Thomas D
Matthews, Keith R
author_facet Briggs, Emma M
Marques, Catarina A
Oldrieve, Guy R
Hu, Jihua
Otto, Thomas D
Matthews, Keith R
author_sort Briggs, Emma M
collection PubMed
description 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 Trypanosoma brucei without prior cell sorting or synchronisation. Additionally, we describe an efficient freeze–thawing protocol that allows single-cell transcriptomic analysis of cryopreserved T. brucei. 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.
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spelling pubmed-102125632023-05-26 Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single-cell transcriptomics Briggs, Emma M Marques, Catarina A Oldrieve, Guy R Hu, Jihua Otto, Thomas D Matthews, Keith R eLife Cell Biology 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 Trypanosoma brucei without prior cell sorting or synchronisation. Additionally, we describe an efficient freeze–thawing protocol that allows single-cell transcriptomic analysis of cryopreserved T. brucei. 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. eLife Sciences Publications, Ltd 2023-05-11 /pmc/articles/PMC10212563/ /pubmed/37166108 http://dx.doi.org/10.7554/eLife.86325 Text en © 2023, Briggs et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Cell Biology
Briggs, Emma M
Marques, Catarina A
Oldrieve, Guy R
Hu, Jihua
Otto, Thomas D
Matthews, Keith R
Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single-cell transcriptomics
title Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single-cell transcriptomics
title_full Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single-cell transcriptomics
title_fullStr Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single-cell transcriptomics
title_full_unstemmed Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single-cell transcriptomics
title_short Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single-cell transcriptomics
title_sort profiling the bloodstream form and procyclic form trypanosoma brucei cell cycle using single-cell transcriptomics
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10212563/
https://www.ncbi.nlm.nih.gov/pubmed/37166108
http://dx.doi.org/10.7554/eLife.86325
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