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Expansion microsopy reveals Plasmodium falciparum blood-stage parasites undergo anaphase with a chromatin bridge in the absence of mini-chromosome maintenance complex binding protein

View ORCID ProfileBenjamin Liffner, View ORCID ProfileSabrina Absalon
doi: https://doi.org/10.1101/2021.09.25.461816
Benjamin Liffner
1Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, IN, USA
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Sabrina Absalon
1Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, IN, USA
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  • For correspondence: Sabsalon@iu.edu
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ABSTRACT

Mitosis in the malaria parasite Plasmodium falciparum undergoes closed mitosis, which occurs within an intact nuclear envelope, and differs significantly from its human host. Mitosis is underpinned by the dynamics of microtubules and the nuclear envelope. To date, our ability to study P. falciparum mitosis by microscopy has been hindered by the small size of P. falciparum nuclei. Ultrastructure expansion microscopy (U-ExM) has recently been developed for P. falciparum, allowing visualization of mitosis at the individual nucleus level. Using U-ExM, three intranuclear microtubule structures are observed: hemispindles, mitotic spindles and interpolar spindles. A previous study demonstrated that the mini-chromosome maintenance complex binding-protein (MCMBP) depletion caused abnormal nuclear morphology and microtubule defects. To investigate the role of microtubules following MCMBP depletion and study the nuclear envelope in these parasites, we developed the first nuclear stain enabled by U-ExM in P. falciparum. MCMBP deficient parasites show aberrant hemispindles and mitotic spindles. Moreover, anaphase chromatin bridges, and individual nuclei containing multiple microtubule structures were observed following MCMBP knockdown. Collectively, this study refines our model for the phenotype of MCMBP-deficient parasites and highlights the utility of U-ExM coupled with a nuclear envelope stain for studying mitosis in P. falciparum.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted September 26, 2021.
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Expansion microsopy reveals Plasmodium falciparum blood-stage parasites undergo anaphase with a chromatin bridge in the absence of mini-chromosome maintenance complex binding protein
Benjamin Liffner, Sabrina Absalon
bioRxiv 2021.09.25.461816; doi: https://doi.org/10.1101/2021.09.25.461816
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Expansion microsopy reveals Plasmodium falciparum blood-stage parasites undergo anaphase with a chromatin bridge in the absence of mini-chromosome maintenance complex binding protein
Benjamin Liffner, Sabrina Absalon
bioRxiv 2021.09.25.461816; doi: https://doi.org/10.1101/2021.09.25.461816

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