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1.
BMC Biol ; 19(1): 69, 2021 04 23.
Article in English | MEDLINE | ID: mdl-33888092

ABSTRACT

BACKGROUND: Rodent malaria parasites (RMPs) serve as tractable tools to study malaria parasite biology and host-parasite-vector interactions. Among the four RMPs originally collected from wild thicket rats in sub-Saharan Central Africa and adapted to laboratory mice, Plasmodium vinckei is the most geographically widespread with isolates collected from five separate locations. However, there is a lack of extensive phenotype and genotype data associated with this species, thus hindering its use in experimental studies. RESULTS: We have generated a comprehensive genetic resource for P. vinckei comprising of five reference-quality genomes, one for each of its subspecies, blood-stage RNA sequencing data for five P. vinckei isolates, and genotypes and growth phenotypes for ten isolates. Additionally, we sequenced seven isolates of the RMP species Plasmodium chabaudi and Plasmodium yoelii, thus extending genotypic information for four additional subspecies enabling a re-evaluation of the genotypic diversity and evolutionary history of RMPs. The five subspecies of P. vinckei have diverged widely from their common ancestor and have undergone large-scale genome rearrangements. Comparing P. vinckei genotypes reveals region-specific selection pressures particularly on genes involved in mosquito transmission. Using phylogenetic analyses, we show that RMP multigene families have evolved differently across the vinckei and berghei groups of RMPs and that family-specific expansions in P. chabaudi and P. vinckei occurred in the common vinckei group ancestor prior to speciation. The erythrocyte membrane antigen 1 and fam-c families in particular show considerable expansions among the lowland forest-dwelling P. vinckei parasites. The subspecies from the highland forests of Katanga, P. v. vinckei, has a uniquely smaller genome, a reduced multigene family repertoire and is also amenable to transfection making it an ideal parasite for reverse genetics. We also show that P. vinckei parasites are amenable to genetic crosses. CONCLUSIONS: Plasmodium vinckei isolates display a large degree of phenotypic and genotypic diversity and could serve as a resource to study parasite virulence and immunogenicity. Inclusion of P. vinckei genomes provide new insights into the evolution of RMPs and their multigene families. Amenability to genetic crossing and transfection make them also suitable for classical and functional genetics to study Plasmodium biology.


Subject(s)
Genome , Malaria , Plasmodium , Animals , Democratic Republic of the Congo , Malaria/genetics , Mice , Phylogeny , Plasmodium/genetics , Rats
2.
Malar J ; 18(1): 26, 2019 Jan 25.
Article in English | MEDLINE | ID: mdl-30683099

ABSTRACT

BACKGROUND: The transcriptional regulation that occurs in malaria parasites during the erythrocytic stages of infection can be studied in vivo with rodent malaria parasites propagated in mice. Time-series transcriptome profiling commonly involves the euthanasia of groups of mice at specific time points followed by the extraction of parasite RNA from whole blood samples. Current methodologies for parasite RNA extraction involve several steps and when multiple time points are profiled, these protocols are laborious, time-consuming, and require the euthanization of large cohorts of mice. RESULTS: A simplified protocol has been designed for parasite RNA extraction from blood volumes as low as 20 µL (microsamples), serially bled from mice via tail snips and directly lysed with TRIzol reagent. Gene expression data derived from microsampling using RNA-seq were closely matched to those derived from larger volumes of leucocyte-depleted and saponin-treated blood obtained from euthanized mice with high reproducibility between biological replicates. Transcriptome profiling of microsamples taken at different time points during the intra-erythrocytic developmental cycle of the rodent malaria parasite Plasmodium vinckei revealed the transcriptional cascade commonly observed in malaria parasites. CONCLUSIONS: Microsampling is a quick, robust and cost-efficient approach to sample collection for in vivo time-series transcriptomic studies in rodent malaria parasites.


Subject(s)
Blood/parasitology , Erythrocytes/parasitology , Gene Expression Profiling/methods , Plasmodium/isolation & purification , RNA, Protozoan/analysis , Animals , Female , Gene Expression Profiling/economics , Gene Expression Profiling/instrumentation , Malaria/blood , Malaria/parasitology , Mice , Mice, Inbred CBA , Plasmodium chabaudi/isolation & purification , Reproducibility of Results
3.
Parasitol Int ; 92: 102680, 2023 Feb.
Article in English | MEDLINE | ID: mdl-36122687

ABSTRACT

Rodent malaria parasites (RMPs) allow the study of malaria parasite biology across its entire life cycle through a vertebrate host and a mosquito vector under laboratory conditions. Among the four RMPs originally collected from wild thicket rats in sub-Saharan Central Africa and adapted to laboratory mice, Plasmodium vinckei has the largest geographical range and includes the largest number of sub-species, demonstrating its deep genetic diversity. Despite affording the same advantages as other RMP species and additionally displaying a large degree of phenotypic and genotypic diversity, P. vinckei has seen limited use in the laboratory. Here, we review the contribution of P. vinckei to our understanding of malaria and highlight the areas where it could offer an advantage over other RMP species in future studies.


Subject(s)
Malaria , Plasmodium , Mice , Rats , Animals , Rodentia/parasitology , Plasmodium/genetics , Malaria/parasitology , Mosquito Vectors
4.
Parasitol Int ; 91: 102635, 2022 Dec.
Article in English | MEDLINE | ID: mdl-35914699

ABSTRACT

In this abbreviated extract of his memoirs of a life in malaria research, Richard Carter (1945-2021) describes the beginnings of malaria parasite genetics research at Edinburgh university, culminating in the first laboratory genetic cross between two strains of malaria parasites. EDITORS' NOTE: This manuscript is an abridged excerpt from Richard Carter's first volume of memoirs describing his career in malaria research. Both volumes, the first concerning malaria parasite genetics, the second focussing on transmission and transmission blocking, are housed in the Wellcome Collection, London [1,2]. Electronic copies of the unabridged memoirs are available to the interested reader upon request to the Editors. These memoirs began as a series of 'recollections' written to a PhD student in March 2014, and were expanded and revised by the author up to 2018. Apart from abridgement, the text is presented in its original form, preserving the informal style in which it was written.


Subject(s)
Malaria , Parasites , Animals , Crosses, Genetic , Humans , Malaria/parasitology , Rodentia
5.
Parasitol Int ; 91: 102650, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36038058

ABSTRACT

In this abbreviated extract of his memoirs of a life in malaria research, Richard Carter (1945-2021) describes the expansion and genetic characterisation of the rodent malaria parasite collection in Edinburgh, culminating in the description of four species, Plasmodium yoelii, Plasmodium berghei, Plasmodium chabaudi and Plasmodium vinckei, and the separation of these into multiple sub-species. The origins of the use of P. chabaudi for the investigation of the genetic determinants of drug resistance in malaria parasites is discussed.


Subject(s)
Malaria , Parasites , Plasmodium , Animals , Malaria/parasitology , Phylogeny , Plasmodium/genetics , Plasmodium berghei/genetics , Rodentia/parasitology
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