Temporal gene expression during asexual development of the apicomplexan Sarcocystis neurona

Donna Huber on May 29, 2024

Fig 1 Progression of the schizont stages during the intracellular development of Sarcocystis neurona expressing yellow fluorescent protein. Post-invasion, the merozoites convert into schizonts that progressively develop into early-, mid-, and late-schizonts while undergoing a form of asexual reproduction called endopolygeny. In the final step in endopolygeny, the mature schizont forms 64 haploid merozoites fully equipped to egress and invade new host cells. S. neurona, Sn; DAPI-stained host cell nuclei, HCN.
Fig 1 Progression of the schizont stages during the intracellular development of Sarcocystis neurona expressing yellow fluorescent protein. Post-invasion, the merozoites convert into schizonts that progressively develop into early-, mid-, and late-schizonts while undergoing a form of asexual reproduction called endopolygeny. In the final step in endopolygeny, the mature schizont forms 64 haploid merozoites fully equipped to egress and invade new host cells. S. neurona, Sn; DAPI-stained host cell nuclei, HCN.

Asexual replication in the apicomplexan Sarcocystis neurona involves two main developmental stages: the motile extracellular merozoite and the sessile intracellular schizont. Merozoites invade host cells and transform into schizonts that undergo replication via endopolygeny to form multiple (64) daughter merozoites that are invasive to new host cells. Given that the capabilities of the merozoite vary significantly from the schizont, the patterns of transcript levels throughout the asexual lifecycle were determined and compared in this study. RNA-Seq data were generated from extracellular merozoites and four intracellular schizont development time points. Of the 6,938 genes annotated in the S. neurona genome, 6,784 were identified in the transcriptome. Of these, 4,111 genes exhibited significant differential expression between the merozoite and at least one schizont development time point. Transcript levels were significantly higher for 2,338 genes in the merozoite and 1,773 genes in the schizont stages. Included in this list were genes encoding the secretory pathogenesis determinants (SPDs), which encompass the surface antigen and SAG-related sequence (SAG/SRS) and the secretory organelle proteins of the invasive zoite stage (micronemes, rhoptries, and dense granules). As anticipated, many of the S. neurona SPD gene transcripts were abundant in merozoites. However, several SPD transcripts were elevated in intracellular schizonts, suggesting roles unrelated to host cell invasion and the initial establishment of the intracellular niche. The hypothetical genes that are potentially unique to the genus Sarcocystis are of particular interest. Their conserved expression patterns are instructive for future investigations into the possible functions of these putative Sarcocystis-unique genes.

Importance: The genus Sarcocystis is an expansive clade within the Apicomplexa, with the species S. neurona being an important cause of neurological disease in horses. Research to decipher the biology of S. neurona and its host-pathogen interactions can be enhanced by gene expression data. This study has identified conserved apicomplexan orthologs in S. neurona, putative Sarcocystis-unique genes, and gene transcripts abundant in the merozoite and schizont stages. Importantly, we have identified distinct clusters of genes with transcript levels peaking during different intracellular schizont development time points, reflecting active gene expression changes across endopolygeny. Each cluster also has subsets of transcripts with unknown functions, and investigation of these seemingly Sarcocystis-unique transcripts will provide insights into the interesting biology of this parasite genus.

Sriveny Dangoudoubiyam, Jamie K Norris, Sivaranjani Namasivayam, Rodrigo de Paula Baptista, Naila Cannes do Nascimento, Joseph Camp, Christopher L Schardl, Jessica C Kissinger, Daniel K Howe. mSphere. 2024 May 29:e0011124. doi: 10.1128/msphere.00111-24.