Dataset Information

Senescent Eimeria acervulina oocysts maintain transcriptional activity during extended refrigerated storage and differerntially express characteristic genes

ABSTRACT: Enteric coccidian parasites harm agriculture and human health. Infectious, sporulated parasites eventually senesce. Here, we examined transcriptional changes in sporulated oocysts of Eimeria acervulina stored for 4-30 months at 4°C. Precipitous decline in RNA abundance and transcription followed an interval of stability. Sixty constitutively expressed genes each contributed >1,000 transcripts per million (TPM) throughout, including a serine protease inhibitor, surface antigen genes, a cation-transporting ATPase, an oocyst wall protein, a zinc finger DHHC domain-containing protein, and highly expressed hypothetical proteins with no known function. Strikingly, ~82% of 6,867 annotated genes underwent differential expression when comparing freshly sporulated parasites to those held for 30 months; nearly one-third of these underwent significant expression change. In freshly sporulated oocysts, 86 significantly DEGs exceeded 1,000 TPM; these encoded heat shock proteins, lactate dehydrogenase, glucose- 6 isomerase, and various hypothetical proteins. The oldest parasites expressed 66 DEGs, including many ribosomal subunits, a haloacid dehalogenase-like hydrolase domain-containing protein, and various hypothetical proteins. Taken together, we identified markers of mature parasites that remain relatively abundant in the transcript pool as oocysts age and identified other transcripts (e.g. ribosomal RNA) that increase in their relative abundance even as RNA abundance declines in senescent parasites.

ORGANISM(S): Eimeria acervulina

PROVIDER: GSE306186 | GEO | 2026/06/03

REPOSITORIES: GEO

ACCESS DATA

Similar Datasets

Project description:Purpose: Parasites of the genus Eimeria, and the closely related human pathogen Cyclospora cayetanensis, cause enteric disease in livestock and people worldwide. These coccidian parasites must undergo maturation (sporulation) before becoming infectious. Understanding oocyst maturation would benefit attempts to reduce the harm they cause to veterinary and human health. No animal model exists to study C. cayetanensis and the parasite cannot be grown in culture. We assessed transcription in maturing oocysts of Eimeria acervulina, a widespread chicken parasite, in order to identify markers of viability and those that may differentiate immature and mature oocysts. We also investigated predicted gene function and determined which of this parasite’s genes also occur in C. cayetanensis. Methods: E. acervulina oocysts were collected from chickens (3 biological replicates) and allowed to sporulate for 24 hours. RNA was extracted from oocysts that were collected every four hours and subjected to RNA-Sequencing. Data were screened for constitutively expressed genes throughout the time course. We also identified differentially expressed genes between immature and mature oocysts. We performed functional characterization of constitutively -expressed and significantly differentially-expressed genes and also biochemical pathway analysis. RT–qPCR validation on selected constitutively and differentially-expressed genes was performed using SYBR Green assays. Results: RNA-Seq yielded ~2 billion paired-end reads, 92% of which mapped to the E. acervulina genome. The ~6,900 annotated genes underwent temporally-coordinated patterns of gene expression. Fifty-three constitutively expressed genes each contributed >1,000 TPM throughout, including those encoding cation-transporting ATPases, an oocyst wall protein, a palmitoyltransferase, membrane proteins, and hypothetical proteins. These genes were enriched for 285 gene ontology (GO) terms and 13 genes were associated with a total of 17 KEGG pathways, defining housekeeping processes and functions important throughout sporulation. Expression of more than 40% of these genes (2,928) differed in sporulated vs. unsporulated oocysts; of these, nearly two-thirds (1,843) increased their expression over time. Eight genes expressed most in unsporulated oocysts, and encoding proteins promoting oocyst maturation and development, could be assigned to 37 GO terms and 5 KEGG pathways. Fifty-six genes underwent significant upregulation in sporulated oocysts, each contributing at least 1,000 TPM. Of these, 40 were annotated by 215 GO assignments and 9 were associated with 18 KEGG pathways, encoding products involved in respiration, carbon fixation, energy utilization, invasion, motility, and stress and detoxification responses. Several RT-qPCR assays confirmed RNA-Seq as a valid means to estimate gene expression. Conclusions: Sporulation orchestrates coordinated changes in the expression of many coccidian genes, most especially those important to metabolic activity. These data elucidate the biology of coccidian development, provide means to evaluate the maturity of parasite cohorts, and may prove useful in predicting infectiousness. Moreover, because many of these genes have homologues in C. cayetanensis, they may prove useful as biomarkers for infectiousness.

Project description:Phosphorylation is a regulatory mechanism by which intracellular apicomplexan parasites control the process of invading and modifying host cells. However, the sporulated oocysts’ phosphoprotein repertoires between virulent and avirulent strains of Toxoplasma gondii and the underlying mechanisms contributing to virulence is still a mystery. A quantitative analysis of the sporulated oocysts’ phosphoproteome profile of T. gondii virulent and avirulent strains belonging to different genotypes was conducted to reveal phosphoprotein expression patterns that contribute to the virulence of a particular phenotype. Phosphopeptides from the sporulated oocysts of the virulent strain PYS (Chinese ToxoDB#9) and the avirulent strain type II (PRU strain) were enriched by titanium dioxide (TiO2) affinity chromatography and quantified using iBT technology. A total of 10,645 unique phosphopeptides, 8,181 nonredundant phosphorylation sites and 2,792 phosphoproteins were identified. The quantitative analysis identified 4,129 differentially expressed phosphopeptides (DEPs) between sporulated oocysts of the PYS strain and PRU strain (|log1.5 fold change| > 1 and P < 0.05), including 2,485 upregulated and 1,644 downregulated phosphopeptides. Motif analysis identified 24 motifs from the upregulated phosphorylated peptides including 22 serine motifs and two threonine motifs (TPE and TP), and 15 motifs from the downregulated phosphorylated peptides including 12 serine motifs and three threonine motifs (TP, RxxT and KxxT) in PYS strain when comparing PYS/PRU. Several kinases were consistent with motifs of overrepresented phosphopeptides, such as PKA, PKG, CKII, IKK, MAPK, EGFR, INSR, Jak, Syk, Src, Ab1. GO analysis, KEGG pathway analysis and STRING analysis revealed differentially expressed phosphoproteins (DEPs) exhibiting significant functional properties. Kinase associated network analysis showed that AGC kinase had the greatest connected peptides. Our findings reveal significant difference in phosphopeptides of sporulated oocysts between virulent and avirulent T. gondii strains, which provides solid foundation for further exploring the mechanisms contributing to the virulence of T. gondii.

Dataset Information

Senescent Eimeria acervulina oocysts maintain transcriptional activity during extended refrigerated storage and differerntially express characteristic genes

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets