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Bioluminescence Imaging to Detect Late Stage Infection of African Trypanosomiasis
Published on: May 18, 2016
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Revealing spatio-temporal dynamics with long-term trypanosomatid live-cell imaging
Richard S Muniz1, Paul C Campbell1, Thomas E Sladewski1
1Department of Molecular Microbiology and Immunology, Brown University, Providence, Rhode Island, United States of America.
Plos Pathogens
|January 18, 2022
Summary
This study introduces a new live-cell imaging method for Trypanosoma brucei, enabling detailed observation of cell division. The technique reveals asymmetric cell division rates and flagellar detachment dynamics, advancing our understanding of these parasites.
Area of Science:
- Cell Biology
- Parasitology
- Microscopy
Background:
- Trypanosoma brucei causes human African trypanosomiasis, but its motility complicates live-cell imaging, hindering research.
- Understanding T. brucei cell division and dynamics is crucial for developing effective treatments.
Purpose of the Study:
- To develop a novel live-cell imaging method for T. brucei to overcome previous limitations.
- To investigate cell division dynamics, daughter cell division rates, and flagellar phenotypes in T. brucei.
Main Methods:
- Developed an agarose microwell system for long-term (24h) confinement and imaging of live T. brucei.
- Utilized "sentinel" cells to monitor imaging toxicity during experiments like RNAi and small-molecule inhibition.
- Applied high-resolution live-cell imaging to track individual cells through multiple divisions.
Main Results:
- Demonstrated that asymmetric daughter cells in T. brucei divide at different rates, with the old-flagellum cell dividing first.
- Observed stepwise flagellar detachment during TbPLK inhibition, with the new flagellum initially tip-linked to the old.
- Found that cells initiating a proposed "back-up" cytokinesis mechanism do not appear to complete division.
Conclusions:
- The developed microwell imaging method enables unprecedented long-term, high-resolution live-cell studies of T. brucei.
- Revealed novel insights into T. brucei cell division asymmetry, flagellar dynamics, and cytokinesis.
- This method opens new avenues for studying previously inaccessible cellular events in trypanosomatids.
