Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294.

Winzer, Pablo Arnold; Anghel, Nicoleta; Imhof, Dennis; Balmer, Vreni; Ortega-Mora, Luis-Miguel; Ojo, Kayode K; Van Voorhis, Wesley C; Müller, Joachim; Hemphill, Andrew (2020). Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294. Pathogens, 9(5) MDPI AG 10.3390/pathogens9050382

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BACKGROUND

Bumped kinase inhibitors (BKIs) are potential drugs for neosporosis treatment in farm animals. BKI-1294 exposure results in the formation of multinucleated complexes (MNCs), which remain viable in vitro under constant drug pressure. We investigated the formation of BKI-1294 induced MNCs, the re-emergence of viable tachyzoites following drug removal, and the localization of CDPK1, the molecular target of BKIs.

METHODS

N. caninum tachyzoites and MNCs were studied by TEM and immunofluorescence using antibodies directed against CDPK1, and against NcSAG1 and IMC1 as markers for tachyzoites and newly formed zoites, respectively.

RESULTS

After six days of drug exposure, MNCs lacked SAG1 surface expression but remained intracellular, and formed numerous zoites incapable of disjoining from each other. Following drug removal, proliferation continued, and zoites lacking NcSAG1 emerged from the periphery of these complexes, forming infective tachyzoites after 10 days. In intracellular tachyzoites, CDPK1 was evenly distributed but shifted towards the apical part once parasites were extracellular. This shift was not affected by BKI-1294.

CONCLUSIONS

CDPK1 has a dynamic distribution depending on whether parasites are located within a host cell or outside. During MNC-to-tachyzoite reconversion newly formed tachyzoites are generated directly from MNCs through zoites of unknown surface antigen composition. Further in vivo studies are needed to determine if MNCs could lead to a persistent reservoir of infection after BKI treatment.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Parasitology

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Winzer, Pablo Arnold; Anghel, Nicoleta; Imhof, Dennis; Balmer, Vreni; Müller, Joachim and Hemphill, Andrew

Subjects:

600 Technology > 630 Agriculture

ISSN:

2076-0817

Publisher:

MDPI AG

Language:

English

Submitter:

Andrew Hemphill

Date Deposited:

17 Feb 2021 14:06

Last Modified:

17 Feb 2021 14:06

Publisher DOI:

10.3390/pathogens9050382

PubMed ID:

32429314

Uncontrolled Keywords:

bumped kinase inhibitor calcium-dependent protein kinase immunofluorescence immunogold labeling inner membrane complex neosporosis surface antigen tachyzoite transmission electron microscopy

BORIS DOI:

10.48350/152372

URI:

https://boris.unibe.ch/id/eprint/152372

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