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Motility and cytoskeletal organisation in the archigregarine Selenidium pygospionis (Apicomplexa): observations on native and experimentally affected parasites

Abstract

Representatives of Apicomplexa perform various kinds of movements that are linked to the different stages of their life cycle. Ancestral apicomplexan lineages, including gregarines, represent organisms suitable for research into the evolution and diversification of motility within the group. The vermiform trophozoites and gamonts of the archigregarine Selenidium pygospionis perform a very active type of bending motility. Experimental assays and subsequent light, electron, and confocal microscopic analyses demonstrated the fundamental role of the cytoskeletal proteins actin and tubulin in S. pygospionis motility and allowed us to compare the mechanism of its movement to the gliding machinery (the so-called glideosome concept) described in apicomplexan zoites. Actin-modifying drugs caused a reduction in the movement speed (cytochalasin D) or stopped the motility of archigregarines completely (jasplakinolide). Microtubule-disrupting drugs (oryzalin and colchicine) had an even more noticeable effect on archigregarine motility. The fading and disappearance of microtubules were documented in ultrathin sections, along with the formation of α-tubulin clusters visible after the immunofluorescent labelling of drug-treated archigregarines. The obtained data indicate that subpellicular microtubules most likely constitute the main motor structure involved in S. pygospionis bending motility, while actin has rather a supportive function.

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Abbreviations

CLSM:

confocal laser scanning microscopy

COL:

colchicine

CYT D:

cytochalasin D

DMSO:

dimethyl sulfoxide

FITC:

fluorescein isothiocyanate

IFA:

indirect immunofluorescent assay

IMC:

inner membrane complex

JAS:

jasplakinolide

LM:

light microscopy

ORY:

oryzalin

PBS:

phosphate-buffered saline

PFA:

4% paraformaldehyde

RR:

ruthenium red

SEM:

scanning electron microscopy

SW:

seawater

TEM:

transmission electron microscopy

TRITC:

tetramethylrhodamine isothiocyanate

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Acknowledgements

Financial support for MK, AV, and AD was provided by the Czech Science Foundation, project No. GBP505/12/G112 (ECIP - Centre of excellence). NV received financial support from MEYS CR (LO1212) and EC (CZ.1.05/2.1.00/01.0017); GGP, from St. Petersburg State University (Grants 1.42.1493.2015, 1.42.1099.2016, project 109-9017); GGP and TGS, from the Russian Foundation for Basic Research (Grant 18-04-00324). We are grateful to Prof. Dominique Soldati-Favre (University of Geneva) for providing the monoclonal anti-actin antibody.

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Correspondence to Magdaléna Kováčiková.

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Andrei Diakin was deceased on 5 November 2018

Electronic supplementary material

Online Resource 1

Bending motility of Selenidium pygospionis incubated in seawater. (MP4 1730 kb)

Online Resource 2

Modified motility of Selenidium pygospionis after treatment with 10 μm oryzalin for 180 min. (MP4 6291 kb)

Online Resource 3

Modified motility of Selenidium pygospionis after treatment with 100 mM colchicine for 40–50 min. (MP4 4240 kb)

Online Resource 4

Modified motility of Selenidium pygospionis after treatment with 30 μm JAS for 150–180 min. (MP4 4311 kb)

Online Resource 5

Modified motility of Selenidium pygospionis after treatment with 30 μm cytochalasin D for 120 min. (MP4 2282 kb)

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Kováčiková, M., Paskerova, G.G., Diakin, A. et al. Motility and cytoskeletal organisation in the archigregarine Selenidium pygospionis (Apicomplexa): observations on native and experimentally affected parasites. Parasitol Res 118, 2651–2667 (2019). https://doi.org/10.1007/s00436-019-06381-z

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Keywords

  • Actin
  • Cytoskeletal drugs
  • Microtubules
  • Motility
  • Ultrastructure
  • α-Tubulin