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

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Summary

African trypanosomes have distinct life cycle stages, but the regulation of their cell replication is poorly understood. This study reveals cell cycle regulated transcriptomes for both forms, identifying conserved and unique gene expression patterns.

Keywords:
Trypanosoma bruceicell biologycell cycleinfectious diseasemicrobiologysingle-cell transcriptomics

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Area of Science:

  • Molecular Biology
  • Parasitology
  • Genomics

Background:

  • African trypanosomes exist as bloodstream forms (BSFs) in mammals and procyclic forms in tsetse flies, crucial for host colonization and life cycle progression.
  • Mechanisms governing the cell replication cycle of these distinct trypanosome forms remain largely unelucidated.

Purpose of the Study:

  • To characterize the cell cycle-regulated (CCR) transcriptomes of both procyclic and slender BSF *Trypanosoma brucei* using single-cell transcriptomics.
  • To develop and validate an efficient protocol for single-cell transcriptomic analysis of cryopreserved *T. brucei*.
  • To computationally reconstruct cell cycle dynamics and compare gene expression patterns between life cycle forms.

Main Methods:

  • Single-cell RNA sequencing (scRNA-seq) applied to unsynchronized *T. brucei* populations.
  • Development of a freeze-thawing protocol for cryopreserved trypanosomes compatible with scRNA-seq.
  • Computational analysis using periodic pseudotime inference to reconstruct cell cycle dynamics.
  • Comparative transcriptomics and proteomic analysis.

Main Results:

  • High-resolution CCR transcriptomes were obtained for both procyclic and slender BSF *T. brucei* without cell synchronization.
  • A core cycling transcriptome conserved across both life cycle forms was identified, alongside form-specific dynamic gene expression.
  • A delay between peak transcript and protein expression was observed for most cycling genes.

Conclusions:

  • This study provides novel insights into the CCR transcriptomes of *T. brucei* life cycle forms.
  • The findings highlight conserved and divergent gene regulation strategies between mammalian and insect stages.
  • An interactive webtool is available for further exploration of the generated transcriptomic data.