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

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,...

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Authors: Winzer, Pablo, Anghel, Nicoleta, Imhof, Dennis, Balmer, Vreni, Ortega Mora, Luis Miguel, Ojo, Kayode K., Van Voorhis, Wesley C., Müller, Joachim, Hemphill, Andrew
Format: article
Publication Date:2020
Country:España
Institution:Universidad Complutense de Madrid (UCM)
Repository:Docta Complutense
Language:English
OAI Identifier:oai:docta.ucm.es:20.500.14352/8055
Online Access:https://hdl.handle.net/20.500.14352/8055
Access Level:Open access
Keyword:bumped kinase inhibitor
calcium-dependent protein kinase
immunofluorescence
immunogold labeling
inner membrane complex
neosporosis
surface antigen
tachyzoite
transmission electron microscopy
Microbiología (Veterinaria)
Farmacología veterinaria
3109.05 Microbiología
3109.08 Farmacología
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oai_identifier_str oai:docta.ucm.es:20.500.14352/8055
network_acronym_str ES
network_name_str España
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spelling Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294Winzer, PabloAnghel, NicoletaImhof, DennisBalmer, VreniOrtega Mora, Luis MiguelOjo, Kayode K.Van Voorhis, Wesley C.Müller, JoachimHemphill, Andrewbumped kinase inhibitorcalcium-dependent protein kinaseimmunofluorescenceimmunogold labelinginner membrane complexneosporosissurface antigentachyzoitetransmission electron microscopyMicrobiología (Veterinaria)Farmacología veterinaria3109.05 Microbiología3109.08 FarmacologíaBackground: 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.MDPIUniversidad Complutense de Madrid20202020-05-1620202020-05-16journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.14352/8055reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Atribución 3.0 Españahttps://creativecommons.org/licenses/by/3.0/es/info:eu-repo/semantics/openAccessoai:docta.ucm.es:20.500.14352/80552026-06-02T12:44:21Z
dc.title.none.fl_str_mv Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
spellingShingle Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
Winzer, Pablo
bumped kinase inhibitor
calcium-dependent protein kinase
immunofluorescence
immunogold labeling
inner membrane complex
neosporosis
surface antigen
tachyzoite
transmission electron microscopy
Microbiología (Veterinaria)
Farmacología veterinaria
3109.05 Microbiología
3109.08 Farmacología
title_short Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_full Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_fullStr Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_full_unstemmed Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
title_sort Neospora caninum: Structure and Fate of Multinucleated Complexes Induced by the Bumped Kinase Inhibitor BKI-1294
dc.creator.none.fl_str_mv Winzer, Pablo
Anghel, Nicoleta
Imhof, Dennis
Balmer, Vreni
Ortega Mora, Luis Miguel
Ojo, Kayode K.
Van Voorhis, Wesley C.
Müller, Joachim
Hemphill, Andrew
author Winzer, Pablo
author_facet Winzer, Pablo
Anghel, Nicoleta
Imhof, Dennis
Balmer, Vreni
Ortega Mora, Luis Miguel
Ojo, Kayode K.
Van Voorhis, Wesley C.
Müller, Joachim
Hemphill, Andrew
author_role author
author2 Anghel, Nicoleta
Imhof, Dennis
Balmer, Vreni
Ortega Mora, Luis Miguel
Ojo, Kayode K.
Van Voorhis, Wesley C.
Müller, Joachim
Hemphill, Andrew
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidad Complutense de Madrid
dc.subject.none.fl_str_mv bumped kinase inhibitor
calcium-dependent protein kinase
immunofluorescence
immunogold labeling
inner membrane complex
neosporosis
surface antigen
tachyzoite
transmission electron microscopy
Microbiología (Veterinaria)
Farmacología veterinaria
3109.05 Microbiología
3109.08 Farmacología
topic bumped kinase inhibitor
calcium-dependent protein kinase
immunofluorescence
immunogold labeling
inner membrane complex
neosporosis
surface antigen
tachyzoite
transmission electron microscopy
Microbiología (Veterinaria)
Farmacología veterinaria
3109.05 Microbiología
3109.08 Farmacología
description 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.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-05-16
2020
2020-05-16
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.14352/8055
url https://hdl.handle.net/20.500.14352/8055
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 3.0 España
https://creativecommons.org/licenses/by/3.0/es/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Atribución 3.0 España
https://creativecommons.org/licenses/by/3.0/es/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Docta Complutense
instname:Universidad Complutense de Madrid (UCM)
instname_str Universidad Complutense de Madrid (UCM)
reponame_str Docta Complutense
collection Docta Complutense
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869411726999420928
score 15.300719

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