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Global expression profiling reveals shared and distinct transcript signatures in arrested act2(-) and CDPK4(-) Plasmodium berghei gametocytes

ABSTRACT: Gametocytogenesis and gametogenesis in malaria parasites are complex processes of cell differentiation and development likely involving many gene products. Gametocytes develop in the blood of the vertebrate host but mature gametocytes are not activated until taken up by the mosquito vector. Several distinct mutants have been described that block gametogenesis but the detailed molecular causes for the mutant phenotypes are not understood. To investigate whether a block in gametogenesis also results in a changed transcriptional profile, we studied two gene deletions mutants; act2(-) lacking stage-specific actin II and CDPK4(-) lacking calcium-dependent protein kinase 4. Whole-genome microarray analysis was performed from RNA of mature gametocytes to compare the transcriptomes of the mutants with wild-type Plasmodium berghei. The microarray analysis identified ~12% of all genes being differentially expressed in either or both mutants compared to normal gametocytes, as defined by at least two-fold change in transcript abundance. A large proportion of differentially expressed genes in both mutants overlapped consistent with the developmental gametocyte arrest. Distinct profiles in each mutant were also observed.

ORGANISM(S): Plasmodium berghei

PROVIDER: GSE65032 | GEO | 2015/09/15

SECONDARY ACCESSION(S): PRJNA272738

REPOSITORIES: GEO

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Global expression profiling reveals shared and distinct transcript signatures in arrested act2(-) and CDPK4(-) Plasmodium berghei gametocytes

Project description:Gametocytogenesis and gametogenesis in malaria parasites are complex processes of cell differentiation and development likely involving many gene products. Gametocytes develop in the blood of the vertebrate host but mature gametocytes are not activated until taken up by the mosquito vector. Several distinct mutants have been described that block gametogenesis but the detailed molecular causes for the mutant phenotypes are not understood. To investigate whether a block in gametogenesis also results in a changed transcriptional profile, we studied two gene deletions mutants; act2(-) lacking stage-specific actin II and CDPK4(-) lacking calcium-dependent protein kinase 4. Whole-genome microarray analysis was performed from RNA of mature gametocytes to compare the transcriptomes of the mutants with wild-type Plasmodium berghei. The microarray analysis identified ~12% of all genes being differentially expressed in either or both mutants compared to normal gametocytes, as defined by at least two-fold change in transcript abundance. A large proportion of differentially expressed genes in both mutants overlapped consistent with the developmental gametocyte arrest. Distinct profiles in each mutant were also observed. Microarray experiments were performed as dual-color hybridizations on Agilent-024169 custom whole genome Plasmodium berghei 44K arrays. To compensate for dye-specific effects, a dye-reversal color-swap was applied.

2015-09-15 | E-GEOD-65032 | biostudies-arrayexpress

Plasmodium berghei transcriptome for female gametocytes, male gametocytes, and asexual erythrocytic stages

Project description:Plasmodium berghei transcriptome for female gametocytes, male gametocytes, and asexual erythrocytic stages

2017-08-21 | GSE102167 | GEO

A G-protein-coupled receptor modulates gametogenesis via PKG-mediated signaling cascade in Plasmodium berghei

Project description:Gametogenesis is essential for malaria parasite transmission, but the molecular mechanism of this process remains to be refined. Here, we identified a G-protein-coupled receptor 180 (GPR180) that plays a critical role in signal transduction during gametogenesis in Plasmodium. In the rodent parasite P. berghei, PbGPR180 was expressed during asexual and sexual development, with more prominent expression in gametocytes and ookinetes. While PbGPR180 was predominantly localized to cytoplasmic puncta in asexual stages and gametocytes, it became associated with the plasma membrane in female gametes and ookinetes. Knockout of pbgpr180 (Δpbgpr180) had no noticeable effect on asexual development or gametocytogenesis but impaired gamete formation and reduced transmission of the parasites to mosquitoes. Transcriptome analysis of the Δpbgpr180 gametocytes revealed a large proportion of the dysregulated genes with assigned functions in cyclic nucleotide signaling transduction, especially the cGMP-PKG-Ca2+ signaling cascade. Deletion of pbgpr180 resulted in a delay and reduction in cytosolic Ca2+ mobilization during the induction of gametogenesis. In addition, the intracellular cGMP level was significantly reduced in the Δpbgpr180 gametocytes, suggesting that PbGPR180 functions upstream of the cGMP-PKG-Ca2+ signaling pathway. In line with this function, PbGPR180 protein was found to interact with several transmembrane transporter proteins and a small GTPase, Rab6, in activated gametocytes. Allele replacement of pbgpr180 with the P. vivax ortholog pvgpr180 showed equal competence of the transgenic parasite in sexual development, suggesting functional conservation of this gene in Plasmodium spp . These results indicate that GPR180 is involved in cGMP-PKG-Ca2+ signal transduction to regulate gametogenesis in malaria parasites.

2022-04-01 | PXD030837 | Pride

PFNF54-Pfs16-GFP-LUC gametocyte time course from commitment to maturity

Project description:Plasmodium falciparum gametocytes undergo 14 days of gametocytogenesis to generate mature, transmissible gametocytes. This timecourse delineates the transcriptional profiles of gametocytes from commitment to maturity.

2018-04-30 | GSE104889 | GEO

Plasmodium berghei Gametocytogenesis

Project description:Transcriptional profiling of gametocyte non-producer lines in Plasmodium berghei Transcriptome of gametocyte non producer lines (natural and genetic KO) and parental (820) lines. The aim of the study was to identify key genes involved in the decision to commit to gametocytogenesis in Plasmodium berghei. These microarrays compare naturally selected lines that do not produce gametocytes, and the parental line and additionally a genetic knock out of AP2-G PBANKA_143750. Data published Sinha, Hughes, et, al Nature tbc. 2- colour microarray comparing to common background pool (containing all life cycle stages). Replicates of different life cycle stages of gametocyte non-producer lines and wild tye (WT) parental control lines

2013-12-13 | E-GEOD-53246 | biostudies-arrayexpress

Plasmodium berghei Gametocytogenesis

2013-12-13 | GSE53246 | GEO

Transcriptome analysis of immature and mature male and female gametocytes of the malaria parasite Plasmodium falciparum

Project description:In the human host, Plasmodium falciparum parasites propagate asexually in erythrocytes causing the symptoms of malaria. However, in every asexual cycle, a small proportion of the parasites commits to sexual differentiation, which is critical for transmission. During a period of about 10 days, the sexually committed parasites either develop into male or female gametocytes. In this study, gametocytes carrying an endogenously GFP tagged version of the female specifically expressed ABCG2 gene were FACS sorted to separate male (GFP low) and female (GFP high) gametocyte populations at different days of development. RNA was subsequently extracted from these populations as well as from asexual parasites as controls and subjected to directional RNAseq. These data reveal the transcriptional differences between asexual parasite stages and developing gametocytes, but also between male and female gametocytes as well as for each sex during the course of differentiation. In particular, male gametocytes undergo distinct transcriptional transitions when comparing the transcriptome between day 4, 6 and 10 of gametocytogenesis. In contrast, female gametocyte transcriptomes are more static during the course of gametocytogenesis and only show minimal variation. This is the first study to longitudinally monitor gene expression in developing male and female gametocytes

2021-07-31 | GSE180985 | GEO

Differential gene expression in ap2-g and ap2-g2 mutants of Plasmodium berghei

Project description:Commitment to and completion of sexual development are essential for malaria parasites to be transmitted through mosquitoes. The molecular mechanism(s) responsible for these processes however, remain largely unknown. We have identified two transcription factors (both belonging to the AP2 family) essential for gametocytogenesis. AP2-G mutants are characterised by a complete inability to produce gametocytes. In AP2-G2 mutants the gametocytaemia is very significantly reduced but not completely abolished. We have performed the microarray experiments in order to cokmpare the transcriptomes of these mutantnts to the WT parasites and between each other. As P.berghei parasites are characterised by asynchronous development in the rodent host, the different stage composition of the sample would impact the analysis. Therefore parasites were harvested and matured in in vitro to the schizont stage Pairwaise comparison between the mutants and parental line was performed. 3 biological replicates of each condition were used.

2013-12-03 | E-GEOD-52859 | biostudies-arrayexpress

Analysis of sex-specific lipid metabolism of Plasmodium falciparum points to the importance of sphingomyelin for gametocytogenesis

Project description:Male and female Plasmodium falciparum gametocytes are the parasite lifecycle stage responsible for transmission of malaria from the human host to the mosquito vector. Not only are gametocytes able to survive in radically different host environments, but they are also precursors for male and female gametes that reproduce sexually soon after ingestion by the mosquito. Here, we investigate the sex-specific lipid metabolism of gametocytes within their host red blood cell. Comparison of the male and female lipidome identifies cholesteryl esters and dihydrosphingomyelin enrichment in female gametocytes. Chemical inhibition of each of these lipid types in mature gametocytes suggests dihydrosphingomyelin synthesis but not cholesteryl ester synthesis is important for gametocyte viability. Genetic disruption of each of the two sphingomyelin synthase genes points towards sphingomyelin synthesis contributing to gametocytogenesis. This study shows that gametocytes are distinct from asexual stages, and that the lipid composition is also vastly different between male and female gametocytes, reflecting the different cellular roles these stages play. Taken together, our results highlight the sex-specific nature of gametocyte lipid metabolism, which has the potential to be targeted to block malaria transmission. This article has an associated First Person interview with the first author of the paper.

2022-04-05 | MTBLS4137 | MetaboLights

Differential gene expression in ap2-g and ap2-g2 mutants of Plasmodium berghei

2013-12-03 | GSE52859 | GEO

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