Previously Unidentified Histone H1-Like Protein Is Involved in Cell Division and Ribosome Biosynthesis in Toxoplasma gondii

Absalon

2023

Molecular Microbiology

Apicomplexan parasites comprise significant pathogens of humans, livestock and wildlife, but also represent a diverse group of eukaryotes with interesting and unique cell biology. The study of cell biology in apicomplexan parasites is complicated by their small size, and historically this has required the application of cutting‐edge microscopy techniques to investigate fundamental processes like mitosis or cell division in these organisms. Recently, a technique called expansion microscopy has been developed, which rather than increasing instrument resolution like most imaging modalities, physically expands a biological sample. In only a few years since its development, a derivative of expansion microscopy known as ultrastructure‐expansion microscopy (U‐ExM) has been widely adopted and proven extremely useful for studying cell biology of Apicomplexa. Here, we review the insights into apicomplexan cell biology that have been enabled through the use of U‐ExM, with a specific focus on Plasmodium, Toxoplasma and Cryptosporidium. Further, we summarize emerging expansion microscopy modifications and modalities and forecast how these may influence the field of parasite cell biology in future.

Section: Potential Future Applications and Emerging Expansion Microsc...mentioning

confidence: 99%

Expansion microscopy of apicomplexan parasites

Absalon

2023

Molecular Microbiology

“…Expansion microscopy is a set of sample preparation techniques that isotropically increase the physical size of a microscopy sample ( Wassie et al, 2019 ). While many expansion microscopy methods have been developed, ultrastructure expansion microscopy (U-ExM) ( Gambarotto et al, 2019 ) was the first used in Plasmodium and since has been used in P. falciparum and across multiple apicomplexan parasites ( Bertiaux et al, 2021 ; Dave et al, 2022 ; Liffner and Absalon, 2021 ; Oliveira Souza et al, 2022 ; Rashpa and Brochet, 2022 ; Severo et al, 2022 ). U-ExM results in the ~4.5-fold isotropic expansion of the sample and largely preserves its proteome, making it compatible with antibody staining and many fluorescent dyes ( Gambarotto et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Atlas of Plasmodium falciparum intraerythrocytic development using expansion microscopy

Diaz

Blauwkamp

et al. 2023

eLife

Apicomplexan parasites exhibit tremendous diversity in much of their fundamental cell biology, but study of these organisms using light microscopy is often hindered by their small size. Ultrastructural expansion microscopy (U-ExM) is a microscopy preparation method that physically expands the sample ∼4.5x. Here, we apply U-ExM to the human malaria parasite Plasmodium falciparum during the asexual blood stage of its lifecycle to understand how this parasite is organized in three-dimensions. Using a combination of dye-conjugated reagents and immunostaining, we have catalogued 13 different P. falciparum structures or organelles across the intraerythrocytic development of this parasite and made multiple observations about fundamental parasite cell biology. We describe that the microtubule organizing center (MTOC) and its associated proteins anchor the nucleus to the parasite plasma membrane during mitosis. Furthermore, the rhoptries, Golgi, basal complex, and inner membrane complex, which form around this anchoring site while nuclei are still dividing, are concurrently segregated and maintain an association to the MTOC until the start of segmentation. We also show that the mitochondrion and apicoplast undergo sequential fission events while maintaining an MTOC association during cytokinesis. Collectively, this study represents the most detailed ultrastructural analysis of P. falciparum during its intraerythrocytic development to date, and sheds light on multiple poorly understood aspects of its organelle biogenesis and fundamental cell biology.

“…Expansion microscopy, which has gained popularity over the last decade, is a set of sample preparation techniques that isotropically increase the physical size of a microscopy sample 2 . While many expansion microscopy methods have been developed, ultrastructure expansion microscopy (U-ExM) 3 was the first used in Plasmodium and since has been used in P. falciparum and across multiple apicomplexan parasites [4][5][6][7][8][9] . U-ExM results in the ~4.5 fold isotropic expansion of the sample and largely preserves its proteome, making it compatible with antibody staining and many fluorescent dyes 3 .…”

Section: Introductionmentioning

confidence: 99%

“…Expansion microscopy is a set of sample preparation techniques that isotropically increase the physical size of a microscopy sample(Wassie, Zhao, & Boyden, 2019). While many expansion microscopy methods have been developed, ultrastructure expansion microscopy (U-ExM) (Gambarotto et al, 2019) was the first used in Plasmodium and since has been used in P. falciparum and across multiple apicomplexan parasites (Bertiaux et al, 2021; Dave, LaFavers, & Arrizabalaga, 2022; Liffner & Absalon, 2021; Oliveira Souza, Jacobs, Back, Bradley, & Arrizabalaga, 2022; R. Rashpa & Brochet, 2022; Severo et al, 2022). U- ExM results in the ∼4.5 fold isotropic expansion of the sample and largely preserves its proteome, making it compatible with antibody staining and many fluorescent dyes (Gambarotto et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Atlas ofPlasmodium falciparumintraerythrocytic development using expansion microscopy

Diaz

Blauwkamp

et al. 2023

Preprint

Apicomplexan parasites exhibit tremendous diversity in much of their fundamental cell biology, but study of these organisms using light microscopy is often hindered by their small size. Ultrastructural expansion microscopy (U-ExM) is a microscopy preparation method that physically expands the sample ~4.5x. Here, we apply U-ExM to the human malaria parasitePlasmodium falciparumduring the asexual blood stage of its lifecycle to understand how this parasite is organized in three-dimensions. Using a combination of dye-conjugated reagents and immunostaining, we have catalogued 13 differentP. falciparumstructures or organelles across the intraerythrocytic development of this parasite and made multiple observations about fundamental parasite cell biology. We describe that the microtubule organizing center (MTOC) and its associated proteins anchor the nucleus to the parasite plasma membrane during mitosis. Furthermore, the rhoptries, Golgi, basal complex, and inner membrane complex, which form around this anchoring site while nuclei are still dividing, are concurrently segregated and maintain an association to the MTOC until the start of segmentation. We also show that the mitochondrion and apicoplast undergo sequential fission events while maintaining an MTOC association during cytokinesis. Collectively, this study represents the most detailed ultrastructural analysis ofP. falciparumduring its intraerythrocytic development to date, and sheds light on multiple poorly understood aspects of its organelle biogenesis and fundamental cell biology.