Germ Plasm Aggregation and Compaction in the Early Zebrafish Embryo

早期斑马鱼胚胎中胚芽质的聚集和压缩

• 批准号: 8784578
• 负责人: Celeste Chloe Eno
• 金额: $ 3.1万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8784578
• 关键词: Actins; Address; Affect; Animals; Auras; Caenorhabditis elegans; Case Study; Cell Communication; Cell Cycle; Cell Differentiation process; Cells; Code; Cytoskeleton; DNA Sequence Rearrangement; Data; Development; Distal; Drosophila genus; Drug usage; Embryo; Embryonic Development; F-Actin; Fertilization; Genes; Germ; Germ Cells; Image; Imagery; In Vitro; Inherited; Injection of therapeutic agent; Intracellular Transport; Laboratories; Lead; Life; Mammals; Messenger RNA; Microfilaments; Microtubules; Modeling; Motor; Movement; Mutation; Myosin ATPase; Myosin Type II; Oocytes; Phenotype; Phylogenetic Analysis; Play; Process; Proteins; Protocols documentation; RNA; Reagent; Reproduction; Research; Role; Slide; Structure; Structure of primordial sex cell; System; Takeda brand of pioglitazone hydrochloride; Technology; Testing; Zebrafish; carcinogenesis; cell determination; cell motility; egg; forward genetics; inhibitor/antagonist; insight; member; mutant; novel; particle; pluripotency; public health relevance; segregation; stemness; teleost

DESCRIPTION (provided by applicant): Germ plasm movement and early acto-myosin movement in the Zebrafish embryo. One of the earliest cell-fate decisions in animal development is the determination of primordial germ cell (PGCs) differentiation, which occurs via inductive cell-cell interactions (e.g. mammals) or preformation (e.g. teleosts, Drosophila, C. elegans). Preformative (maternally inherited) germ plasm is a specialized structure made up of ribonucleoparticles (RNPs), proteins and often associated with cytoskeletal components. The zebrafish model is excellent for studying the movement of germ plasm in early embryogenesis. The transparent embryos are easily manipulated using drugs, as well as morpholino and RNA injections. Recently, members of our laboratory have refined a protocol for manipulation of maternal factors acting in the embryo by injection of reagents into oocytes undergoing in vitro maturation, which has opened the possibility of manipulation prior to and immediately after fertilization. My preliminary data suggest that phosphomyosin is present in germ plasm RNPs during their multimerization prior to and during furrow formation, and that these RNPs reside on F- actin. We propose a mechanism by which myosin II-driven F-actin sliding aggregates germ plasm particles and moves the aggregates into the first two forming furrows. Later, during furrow maturation, germ plasm RNPs continue to undergo multimerization until they form compact masses at the distal ends of these furrows. A maternal effect lethal mutant, aura, does not have the concentric actin rings seen in wild type embryos and do not properly aggregate germ plasm RNPs. This suggests that the protein aura codes for, Mid1ip1L, is involved in the cytoskeletal dynamics of germ plasm early movement. I plan to address these hypotheses by characterizing germ plasm aggregation and recruitment on actin filaments prior to and at furrow initiation, and determining the role Mid1ip1L plays in early cytoskeletal dynamics. The proposed hypothesis of GP RNP movement will provide details on a novel mechanism for the movement of such cellular determinants via the action of a myosin motor on an actin cytoskeletal network. The mechanisms studied will lead to understanding the proper movement of PGC-determining factors, relating to cell stemness, reproduction, carcinogenesis and pluripotency.

描述（由申请人提供）：斑马鱼胚胎中的细菌血浆运动和早期的肌动蛋白运动。动物发育中最早的细胞命运决定之一是确定原始生殖细胞（PGCS）分化，该分化是通过电感细胞 - 细胞相互作用（例如哺乳动物）或预形成（例如Teleotosts，Drosopophila，C。exkelelans）发生的。预向（母体遗传）种植血浆是由核糖核颗粒（RNP），蛋白质，通常与细胞骨架成分相关的专业结构。斑马鱼模型非常适合研究早期胚胎发生中种植血浆的运动。透明的胚胎很容易使用药物以及吗啡和RNA注射来操纵。最近，我们的实验室成员通过将试剂注射到经过体外成熟的卵母细胞中来操纵胚胎的母体因素的方案，该方案在受肥后和立即进行了操纵的可能性。我的初步数据表明，在沟形成之前和期间，在其多聚化期间，磷肌球蛋白存在于种植血浆RNP中，并且这些RNP驻留在F-肌动蛋白上。我们提出了一种机制，通过该机制，肌球蛋白II驱动的F-肌动蛋白滑动聚集蛋白质血浆颗粒并将聚集体移入前两个形成的沟中。后来，在犁沟成熟期间，种植血浆RNP继续进行多聚化，直到它们在这些犁沟的远端形成紧凑的质量为止。母体效应致命突变体（Aura）没有在野生型胚胎中看到的同心肌动蛋白环，也没有正确聚集的种系浆RNP。这表明蛋白Aura代码为MID1IP1L，参与了生殖血浆早期运动的细胞骨架动力学。我计划通过表征在犁沟开始之前和处于肌动蛋白丝上的种植浆聚集和募集来解决这些假设，并确定MID1IP1L在早期细胞骨架动力学中的作用。提出的GP RNP运动的假设将通过肌球蛋白运动在肌动蛋白细胞骨架网络上的作用提供有关这种细胞决定因素运动的新机制的细节。所研究的机制将导致了解与细胞干，繁殖，癌变和多能性有关的PGC决定因素的正确运动。