Developmental Synchrony in Dictyostelium discoideum

这项研究项目旨在调查盘状网柄线虫的发育和同步化，以获得对多细胞生物如何组织自己的有价值的见解。这项研究阐述了在不同水平上理解同步性的更广泛的科学重要性，如细胞、组织和整个生物体，以及它与生物科学的相关性。盘基网柄菌是一种理想的模式生物，因为它的简单和适合于遗传和分子分析。该项目试图揭开细胞协调其发育、组织形成和组装以及生物体在不同发育阶段进行进化的复杂机制。通过采用单细胞RNA测序和标准RNA测序等先进技术，科学家们将量化同步性和基因表达模式。通过将野生型发育与缺乏特定信号机制的突变株进行比较，他们将分析同步遗传和时间扰动的后果。此外，这项研究还将生成一份全面的转录组图谱，确定新的细胞类型和亚型。生成的数据集将向公众开放，允许进一步探索同步和发展。此外，STEM教师将接受数据挖掘和科学研究方面的培训，提高他们的技能并激励他们的学生。这项研究项目有助于提高人们的科学素养，并扩大我们对多细胞生物体发育的复杂生物学过程的理解。本研究项目主要研究盘基网柄菌在细胞、组织和生物体等不同水平上的同步和发育。主要目的是通过检查一系列发育检查点来研究同步性的机制和意义。这些检查点涉及细胞外环磷酸腺苷(CAMP)信号、同种识别蛋白介导的细胞-细胞接触以及孢子分化因子(SDF)多肽的产生。为了实现这些目标，研究人员将使用先进的技术，如单细胞RNA测序(scRNA-seq)和标准RNA测序(RNA-seq)。这些方法将允许量化不同发育阶段的同步性和基因表达模式。分析将涉及比较野生型发育与缺乏胞外cAMP信号的突变株的同步性。此外，还将检查一种基因抑制株，它可以在不恢复同步性的情况下恢复发育，使人们能够更深入地了解同步性扰动的后果。该项目还旨在为盘基网柄菌创建一份全面的单细胞转录组图谱。这份图集将使识别新的细胞类型和亚型成为可能。为了验证这些发现，将通过产生绿色荧光蛋白融合和随后的时空基因表达来进一步研究选定的差异表达基因，并将进行遗传消融实验以探索新鉴定的细胞类型的功能。总体而言，这一研究项目有助于我们理解盘基网柄菌的发育同步性及其潜在的分子机制。先进的分子工具和分析技术的使用将为多细胞生物的协调组织提供有价值的见解。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.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.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.