In Vitro Generation of Plasmodium falciparum Ookinetes

Bounkeua, Viengngeun; Li, Fengwu; Vinetz, Joseph M.

doi:10.4269/ajtmh.2010.10-0433

Cited by 35 publications

(46 citation statements)

References 50 publications

(73 reference statements)

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“…In this method, Karnovsky's fixative and LXR 112 resin were used to prepare the ookinete cells, which resulted in better structural and organelle integrity and resolution than previously observed in Plasmodium (including the incorrect depiction of rhopties in the apical complex) [17][18][19][20][21][22][23][24][25][26][27][28] and Haemoproteus.…”

Section: Resultsmentioning

confidence: 99%

“…Over the past four decades, production of the various species of Plasmodium ookinetes (P. falciparum, 26,37,38 P. vivax, 39 and rodent-infecting parasites including P. berghei, P. yoelii, and P. yoelii nigeriensis [40][41][42] ) in in vitro systems has been a cumbersome procedure and limited by quantities of parasites obtainable by in vitro culture for detailed analysis.…”

Section: Discussionmentioning

confidence: 99%

“…27,34 This extension of apical structure, not previously observed in studies of the fine structure of P. falciparum and P. gallinaceum ookinetes, was previously reported in ookinetes of the rodent-infecting parasite P. yoelii, both in free ookinetes in culture and during penetration of the basement membrane of the mosquito midgut. 26,[48][49][50][51] The Plasmodium life cycle is complex, with sequential, preprogrammed developmental stages in vertebrate and invertebrate hosts. Although the merozoite and sporozoite stage parasites responsible for red cell and liver cell invasion, respectively, contain three distinct organelles-micronemes, rhoptries, and dense granules-the ookinete only seems to have one such organelle, micronemes.…”

Section: Discussionmentioning

confidence: 99%

“…With improved methods for obtaining P. falciparum 26,37,38 and P. vivax ookinetes in vitro, 39,60 studies of these human-infecting parasites are now possible as well. …”

Section: Discussionmentioning

confidence: 99%

“…12 A dense set of variably staining and shaped micronemes, both cigarshaped and rounded, abut microtubules known to be involved in microneme mobilization and secretion. 26 The high-quality visualization of these structures, distinctly seen in Figure 3B, may be the result of improved TEM methods described herein.…”

mentioning

confidence: 92%

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New Ultrastructural Analysis of the Invasive Apparatus of the Plasmodium Ookinete

Patra¹,

Vinetz²

2012

The American Journal of Tropical Medicine and Hygiene

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Abstract. Invasion of the mosquito midgut by the Plasmodium ookinete determines the success of transmission of malaria parasites from humans to mosquitoes and therefore, is a potential target for molecular intervention. Here, we show higher-resolution ultrastructural details of developing and mature P. gallinaceum ookinetes than previously available. Improved fixation and processing methods yielded substantially improved transmission electron micrographs of ookinetes, particularly with regard to visualization of subcellular secretory and other organelles. These new images provide new insights into the synthesis and function of vital invasive machinery focused on the following features: apical membrane protrusions presumptively used for attachment and protein secretion, dark spherical bodies at the apical end of the mature ookinete, and the presence of a dense array of micronemes apposed to microtubules at the apical end of the ookinete involved in constitutive secretion. This work advances understanding of the molecular and cellular details of the Plasmodium ookinete and provides the basis of future, more detailed mechanistic experimentation on the biology of the Plasmodium ookinete.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…With improved methods for obtaining P. falciparum 26,37,38 and P. vivax ookinetes in vitro, 39,60 studies of these human-infecting parasites are now possible as well. …”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 92%

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New Ultrastructural Analysis of the Invasive Apparatus of the Plasmodium Ookinete

Patra¹,

Vinetz²

2012

The American Journal of Tropical Medicine and Hygiene

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A novel flow cytometric application discriminates among the effects of chemical inhibitors on various phases of Babesia divergens intraerythrocytic cycle

et al. 2017

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Human babesiosis is a global emerging infectious disease caused by intraerythrocytic parasites of the genus Babesia. Its biology has remained largely unexplored due to a lack of critical tools and techniques required to define the various stages and phases of the parasite's cycle in its host RBC and the interplay between host and parasite. This article presents a powerful set of tools combining stage synchronization of the parasite with a platform that encompasses both a flow cytometric evaluation of the subpopulation structure of the parasite population together with a morphological assessment of the population parasites using light microscopy of conventional Giemsa stained smears. Together, these yield specific information on the effect of any drug/condition of interest and its targeted biological process, allowing the characterization of the adaptive response of parasites to a particular stressor agent. Three inhibitors were used in this study, each targeting a specific phase of the parasite's lifecycle, neuraminidase for host cell invasion, N-acetyl-L-leucyl-L-leucyl-L-norleucinal for parasite development and EGTA for parasite egress from the host cell. Results presented prove the power of this combination platform in discriminating the specific targets among the life-cycle processes of the parasite-invasion, development/proliferation and egress. This will expand the range of queries that can now be successfully addressed in this parasite, opening avenues for the development of new methods to control babesiosis, either by chemicals (screening for new chemotherapy drugs or defining levels of parasite resistance) or physical methods (light irradiation or heat shock used in pathogen reduction/elimination methods).

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Chemo‐proteomics in antimalarial target identification and engagement

Bailey

Nguyen

Cowman

et al. 2023

Medicinal Research Reviews

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Humans have lived in tenuous battle with malaria over millennia. Today, while much of the world is free of the disease, areas of South America, Asia, and Africa still wage this war with substantial impacts on their social and economic development. The threat of widespread resistance to all currently available antimalarial therapies continues to raise concern. Therefore, it is imperative that novel antimalarial chemotypes be developed to populate the pipeline going forward. Phenotypic screening has been responsible for the majority of the new chemotypes emerging in the past few decades. However, this can result in limited information on the molecular target of these compounds which may serve as an unknown variable complicating their progression into clinical development. Target identification and validation is a process that incorporates techniques from a range of different disciplines. Chemical biology and more specifically chemo‐proteomics have been heavily utilized for this purpose. This review provides an in‐depth summary of the application of chemo‐proteomics in antimalarial development. Here we focus particularly on the methodology, practicalities, merits, and limitations of designing these experiments. Together this provides learnings on the future use of chemo‐proteomics in antimalarial development.

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In Vitro Generation of Plasmodium falciparum Ookinetes

Cited by 35 publications

References 50 publications

New Ultrastructural Analysis of the Invasive Apparatus of the Plasmodium Ookinete

New Ultrastructural Analysis of the Invasive Apparatus of the Plasmodium Ookinete

A novel flow cytometric application discriminates among the effects of chemical inhibitors on various phases of Babesia divergens intraerythrocytic cycle

Chemo‐proteomics in antimalarial target identification and engagement

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