ArticlePloS one2018
A fixable probe for visualizing flagella and plasma membranes of the African trypanosome.
Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 9 citations in OpenAlex.
- A dual-aptazyme system for genetic complementation reveals stage-specific roles for themSphere · 2026Article
- Discovery of effectors for casein kinase signaling in the African trypanosome.Scientific reports · 2025Article
- A limitation lifted: A conditional knockdown system reveals essential roles for Polo-like kinase and Aurora kinase 1 inProceedings of the National Academy of Sciences of the United States of America · 2025Article
- Article
- Solid-Phase Synthesis of Fluorescent Probes for Plasma Membrane Labelling.Molecules (Basel, Switzerland) · 2021Article
- Airyscan Superresolution Microscopy to Study Trypanosomatid Cell Biology.Methods in molecular biology (Clifton, N.J.) · 2020Article
- Kinetoplast Division Factors in a Trypanosome.Trends in parasitology · 2019Review
- Series of Alkynyl-Substituted Thienopyrimidines as Inhibitors of Protozoan Parasite Proliferation.ACS medicinal chemistry letters · 2018Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
The protozoan Trypanosoma brucei sp. cause diseases in humans and animals. Studies of T. brucei cell biology have revealed unique features, such as major endocytic events being limited to a single region, and mitochondrial genome segregation mediated via basal bodies. Further understanding of trypanosome cell biology can be facilitated with super-resolution fluorescence microscopy. Lack of a plasma membrane probe for fixed trypanosomes remains a persistent problem in need of a working solution. Herein, we report protocols developed using mCLING in super-resolution structured illumination fluorescence microscopy (SR-SIM). mCLING comprehensively labels flagellar membranes, including nascent intracellular stages. To extend its usefulness for trypanosome biology we optimized mCLING in combination with organelle-specific antibodies for immunofluorescence of basal bodies or mitochondria. Then in work with live trypanosomes, we demonstrated internalization of mCLING into endocytic stations that overlap with LysoTracker in acidic organelles. Greater detail of the intracellular location of mCLING was obtained with SR-SIM after pulsing trypanosomes with the probe, and allowing continuous uptake of fluorescent concanavalin A (ConA) destined for lysosomes. In most cases, ConA and mCLING vesicles were juxtaposed but not coincident. A video of the complete image stack at the 15 min time point shows zones of mCLING staining surrounding patches of ConA, consistent with persistence of mCLING in membranes of compartments that contain luminal ConA. In summary, these studies establish mCLING as a versatile trypanosome membrane probe compatible with super-resolution microscopy that can be used for detailed analysis of flagellar membrane biogenesis. In addition, mCLING can be used for immunofluorescence in fixed, permeabilized trypanosomes. Its robust staining of the plasma membrane eliminates a need to overlay transmitted light images on fluorescence pictures obtained from widefield, confocal, or super-resolution microscopy.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.