Gene expression in Plasmodium berghei ookinetes and early oocysts in a co-culture system with mosquito cells

Vontas, John; Sidén‐Kiamos, Inga; Papagiannakis, Giorgos; Karras, Marianna; Waters, Andrew P.; Louis, Christos

doi:10.1016/j.molbiopara.2005.01.009

Cited by 7 publications

(7 citation statements)

References 19 publications

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“…pfCDPK3 (PFC420w) was identified in P. falciparum as the third CDPK‐family protein in plasmodium species, and its transcription was reported to occur in the sexual stage (Li et al ., 2000). However, recent EST and DNA microarray studies have showed that P. berghei CDPK3 is transcribed in the ookinete stage (Abraham et al ., 2004; Vontas et al ., 2005). We also analysed our P. berghei EST database and confirmed that six ESTs for PbCDPK3 were included in ookinete ESTs.…”

Section: Resultsmentioning

confidence: 99%

A calcium‐dependent protein kinase regulates Plasmodium ookinete access to the midgut epithelial cell

Ishino¹,

Orito²,

Chinzei³

et al. 2005

Molecular Microbiology

203

187

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SummaryPlasmodium parasites are fertilized in the mosquito midgut and develop into motile zygotes, called ookinetes, which invade the midgut epithelium. Here we show that a calcium-dependent protein kinase, CDPK3, of the rodent malarial parasite ( Plasmodium berghei ) is produced in the ookinete stage and has a critical role in parasite transmission to the mosquito vector. Targeted disruption of the CDPK3 gene decreased ookinete ability to infect the mosquito midgut by nearly two orders of magnitude. Electron microscopic analyses demonstrated that the disruptant ookinetes could not access midgut epithelial cells by traversing the layer covering the cell surface. An in vitro migration assay showed that these ookinetes lack the ability to migrate through an artificial gel, suggesting that this defect caused their failure to access the epithelium. In vitro migration assays also suggested that this motility is induced in the wild type by mobilization of intracellular stored calcium. These results indicate that a signalling pathway involving calcium and CDPK3 regulates ookinete penetration of the layer covering the midgut epithelium. Because humans do not possess CDPK family proteins, CDPK3 is a good target for blocking malarial transmission to the mosquito vector.

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Section: Resultsmentioning

confidence: 99%

A calcium‐dependent protein kinase regulates Plasmodium ookinete access to the midgut epithelial cell

Ishino¹,

Orito²,

Chinzei³

et al. 2005

Molecular Microbiology

203

187

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“…The RNA-seq studies presented here provide information on gene expression at an unprecedented depth and breadth of coverage of multiple blood stages and ookinetes. Previously, only a few large-scale transcriptome (microarray) analyses of P. berghei blood stages and ookinetes had been performed [17,46]. These studies were based on a highly fragmented draft P. berghei genome and, therefore, expression data were only generated for about half of all P. berghei genes.…”

Section: Discussionmentioning

confidence: 99%

A comprehensive evaluation of rodent malaria parasite genomes and gene expression

et al. 2014

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BackgroundRodent malaria parasites (RMP) are used extensively as models of human malaria. Draft RMP genomes have been published for Plasmodium yoelii, P. berghei ANKA (PbA) and P. chabaudi AS (PcAS). Although availability of these genomes made a significant impact on recent malaria research, these genomes were highly fragmented and were annotated with little manual curation. The fragmented nature of the genomes has hampered genome wide analysis of Plasmodium gene regulation and function.ResultsWe have greatly improved the genome assemblies of PbA and PcAS, newly sequenced the virulent parasite P. yoelii YM genome, sequenced additional RMP isolates/lines and have characterized genotypic diversity within RMP species. We have produced RNA-seq data and utilised it to improve gene-model prediction and to provide quantitative, genome-wide, data on gene expression. Comparison of the RMP genomes with the genome of the human malaria parasite P. falciparum and RNA-seq mapping permitted gene annotation at base-pair resolution. Full-length chromosomal annotation permitted a comprehensive classification of all subtelomeric multigene families including the ‘Plasmodium interspersed repeat genes’ (pir). Phylogenetic classification of the pir family, combined with pir expression patterns, indicates functional diversification within this family.ConclusionsComplete RMP genomes, RNA-seq and genotypic diversity data are excellent and important resources for gene-function and post-genomic analyses and to better interrogate Plasmodium biology. Genotypic diversity between P. chabaudi isolates makes this species an excellent parasite to study genotype-phenotype relationships. The improved classification of multigene families will enhance studies on the role of (variant) exported proteins in virulence and immune evasion/modulation.Electronic supplementary materialThe online version of this article (doi:10.1186/s12915-014-0086-0) contains supplementary material, which is available to authorized users.

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“…Although Plasmodium encodes a putative p ABA‐synthetase (Triglia and Cowman, 1999), the endogenous p ABA synthesis is obviously rate‐limiting, rendering the oocysts p ABA‐auxotrophs. Two recent expression profiling analyses now pave the way for a detailed analysis of oocyst development by reverse genetics (Srinivasan et al ., 2004; Vontas et al ., 2005). By employing a polymerase chain reaction‐based enrichment procedure, termed suppression subtractive hybridization, several late oocyst transcripts, including MAEBL (apical membrane antigen/erythrocyte binding‐like protein; Kariu et al ., 2002), were isolated (Srinivasan et al ., 2004).…”

Section: Oocyst Differentiationmentioning

confidence: 99%

“…However, when this approach was used to enrich for mid oocyst‐specific transcripts only a single Plasmodium‐ specific sequence was returned, highlighting the current technical limitations to obtain pure parasite material from infected mosquitoes (Srinivasan et al ., 2004). This limitation was partly overcome by expression profiling of in vitro ‐cultured oocysts using microarrays (Vontas et al ., 2005). A combination of biochemical and genetic characterization of these upregulated genes is now required to study their respective role in oocyst development.…”

Section: Oocyst Differentiationmentioning

confidence: 99%

Getting infectious: formation and maturation of Plasmodium sporozoites in the Anopheles vector

Matuschewski

2006

Cell Microbiol

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SummaryResearch on Plasmodium sporozoite biology aims at understanding the developmental program steering the formation of mature infectious sporozoites -the transmission stage of the malaria parasite. The recent identification of genes that are vital for sporozoite egress from oocysts and subsequent targeting and transmigration of the mosquito salivary glands allows the identification of mosquito factors required for life cycle completion. Mature sporozoites appear to be equipped with the entire molecular repertoire for successful transmission and subsequent initiation of liver stage development. Innovative malaria intervention strategies that target the early, non-pathogenic phases of the life cycle will crucially depend on our insights into sporozoite biology and the underlying molecular mechanisms that lead the parasite from the mosquito midgut to the liver.

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Gene expression in Plasmodium berghei ookinetes and early oocysts in a co-culture system with mosquito cells

Cited by 7 publications

References 19 publications

A calcium‐dependent protein kinase regulates Plasmodium ookinete access to the midgut epithelial cell

A calcium‐dependent protein kinase regulates Plasmodium ookinete access to the midgut epithelial cell

A comprehensive evaluation of rodent malaria parasite genomes and gene expression

Getting infectious: formation and maturation of Plasmodium sporozoites in the Anopheles vector

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