NSF Award
2319686
This research project aims to investigate the development and synchronization of Dictyostelium discoideum, a social soil amoeba, to gain valuable insights into how multicellular organisms organize themselves. This study addresses the broader scientific importance of understanding synchrony at different levels, such as cells, tissues, and whole organisms, and its relevance to biological science. Dictyostelium discoideum is an ideal model organism due to its simplicity and suitability for genetic and molecular analyses. The project seeks to unravel the intricate mechanisms by which cells coordinate their development, tissues form and assemble, and organisms progress through distinct developmental stages. By employing advanced techniques such as single-cell RNA sequencing and standard RNA sequencing, the scientists will quantify synchrony and gene expression patterns. By comparing wild-type development with mutant strains lacking specific signaling mechanisms, they will analyze the consequences of genetic and temporal perturbations in synchrony. Additionally, the research will generate a comprehensive transcriptome atlas, identifying new cell types and subtypes. The generated data sets will be publicly accessible, allowing for further exploration of synchrony and development. Furthermore, STEM teachers will be trained in data mining and scientific research, enhancing their skills and inspiring their students. This project contributes to scientific literacy and expands our understanding of the intricate biological processes underlying multicellular organism development.<br/><br/>This research project focuses on studying the synchronization and development of Dictyostelium discoideum at various levels: cells, tissues, and organisms. The primary objective is to investigate the mechanisms and significance of synchrony by examining a series of developmental checkpoints. These checkpoints involve extracellular cyclic AMP (cAMP) signaling, cell-cell contact mediated by allorecognition proteins, and the production of Spore Differentiation Factor (SDF) peptides. To achieve these goals, the researchers will employ advanced techniques such as single-cell RNA sequencing (scRNA-seq) and standard RNA sequencing (RNA-seq). These methods will allow for the quantification of synchrony and gene expression patterns during different developmental stages. The analysis will involve comparing the synchrony in wild-type development with that of mutant strains lacking extracellular cAMP signaling. Additionally, a genetic suppressor strain will be examined, which restores development without restoring synchrony, enabling a deeper understanding of the consequences of perturbations in synchrony. The project also aims to create a comprehensive single-cell transcriptome atlas for Dictyostelium discoideum. This atlas will enable the identification of novel cell types and subtypes. To validate the findings, selected differentially expressed genes will be further investigated by generating Green Fluorescent Protein fusions and following spatiotemporal gene expression, and genetic ablation experiments will be conducted to explore the functions of newly identified cell types. Overall, this research project contributes to our understanding of developmental synchrony in Dictyostelium discoideum and its underlying molecular mechanisms. The use of advanced molecular tools and analysis techniques will provide valuable insights into the coordinated organization of multicellular organisms.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
