Expansion microscopy facilitates quantitative super-resolution studies of cytoskeletal structures in kinetoplastid parasites

0554621 - ÚMG 2022 RIV GB eng J - Článek v odborném periodiku
Gorilák, Peter - Pružincová, Martina - Váchová, Hana - Olsinova, M. - Schmidt Černohorská, Markéta - Varga, Vladimír
Expansion microscopy facilitates quantitative super-resolution studies of cytoskeletal structures in kinetoplastid parasites.
Open Biology. Roč. 11, č. 9 (2021), č. článku 210131. E-ISSN 2046-2441
Grant CEP: GA ČR GC20-23165J; GA MŠMT(CZ) LM2018129; GA MŠMT(CZ) EF18_046/0016045
Výzkumná infrastruktura: Czech-BioImaging II - 90129
Institucionální podpora: RVO:68378050
Klíčová slova: Trypanosoma brucei * Leishmania major * expansion microscopy * microtubule-based cytoskeleton
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 7.000, rok: 2021
Způsob publikování: Open access
https://royalsocietypublishing.org/doi/10.1098/rsob.210131

Expansion microscopy (ExM) has become a powerful super-resolution method in cell biology. It is a simple, yet robust approach, which does not require any instrumentation or reagents beyond those present in a standard microscopy facility. In this study, we used kinetoplastid parasites Trypanosoma brucei and Leishmania major, which possess a complex, yet well-defined microtubule-based cytoskeleton, to demonstrate that this method recapitulates faithfully morphology of structures as previously revealed by a combination of sophisticated electron microscopy (EM) approaches. Importantly, we also show that due to the rapidness of image acquisition and three-dimensional reconstruction of cellular volumes ExM is capable of complementing EM approaches by providing more quantitative data. This is demonstrated on examples of less well-appreciated microtubule structures, such as the neck microtubule of T. brucei or the pocket, cytosolic and multivesicular tubule-associated microtubules of L. major. We further demonstrate that ExM enables identifying cell types rare in a population, such as cells in mitosis and cytokinesis. Three-dimensional reconstruction of an entire volume of these cells provided details on the morphology of the mitotic spindle and the cleavage furrow. Finally, we show that established antibody markers of major cytoskeletal structures function well in ExM, which together with the ability to visualize proteins tagged with small epitope tags will facilitate studies of the kinetoplastid cytoskeleton.
Trvalý link: http://hdl.handle.net/11104/0329317