Ion signaling and cell state transitions for organ size control of regenerating zebrafish fins

离子信号和细胞状态转变用于控制再生斑马鱼鳍的器官大小

• 批准号: 10631064
• 负责人: Heather K Le Bleu
• 金额: $ 4.65万
• 依托单位: UNIVERSITY OF OREGON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10631064
• 关键词: Adult; Affect; Animals; Back; Bone Diseases; Calcineurin; Calcium; Cardiac; Cell Lineage; Cell Separation; Cells; Cellular biology; Communication; Data; Data Set; Development; Developmental Biology; Disease; Distal; Doctor of Philosophy; Dominant-Negative Mutation; Ectopic Expression; Educational process of instructing; Educational workshop; Event; Genes; Genetic Epistasis; Geometry; Growth; Health; Homologous Gene; Human; Image; Injury; Ion Channel; Ions; Kinetics; Laboratories; Leadership; Long QT Syndrome; Mentors; Mesenchymal; Mesenchyme; Modeling; Molecular; Natural regeneration; Organ; Organ Model; Organ Size; Pathway interactions; Phenotype; Population; Potassium Channel; Production; Proliferating; Publishing; Regenerative Medicine; Regenerative capacity; Reporter; Research; Resected; Role; Science; Shapes; Signal Transduction; Site; Skeleton; Testing; Tissues; Training; Transgenic Organisms; Transplantation; Voltage-Gated Potassium Channel; WNT Signaling Pathway; Woman; Zebrafish; bioelectricity; blastema; bone; calcineurin phosphatase; cell behavior; experience; experimental study; human disease; human tissue; in vivo; injured; mutant; novel strategies; organ repair; prevent; regenerative; regenerative biology; repaired; restoration; skeletal; small molecule; therapeutic target; transcription factor; transcriptomics; voltage

PROJECT SUMMARY This project provides the applicant with Ph.D. training intersecting cell, developmental and regenerative biology. Zebrafish regenerate resected fins, including their bony ray skeletons, back to the original size irrespective of injury extent. Yet, how fins “know” when to stop growing as the correct size is attained is poorly understood. The applicant’s laboratory recently proposed a new model for robust fin size restoration based on initial amount of a skeletal geometry-defined “niche” population and its progressive depletion by a cell state transition. Further, the group found dramatic fin overgrowth in the classic zebrafish mutant longfint2 is caused by cis-ectopic expression of the kcnh2a K+ channel. Kcnh2a in longfint2 results in niche cell perdurance and therefore excessive outgrowth during regeneration. The applicant’s transplant experiments show ectopic kcnh2a in the mesenchyme lineage that generates niche cells is sufficient for overgrowth. Yet, how kcnh2a disrupts orderly niche depletion is unknown. Published studies show inhibiting the Ca2+-dependent phosphatase calcineurin also causes striking fin overgrowth. The applicant’s new epistasis experiments suggest Kcnh2a functions upstream of calcineurin. Their new transgenic Ca2+ reporter identifies dynamic Ca2+ levels in niche/mesenchyme cells that preliminary transcriptomics indicate depend on specifically expressed Ca2+ channels. This leads to the central hypothesis that Ca2+ channel activity and dynamic Ca2+ levels activate calcineurin signaling to promote niche-to-mesenchymal cell state transitions and end fin regrowth. The applicant will pursue three specific aims: 1) Determine how Kcnh2a impacts intracellular Ca2+ dynamics during fin regeneration, 2) Identify Ca2+ channels influencing regenerated fin size, and 3) Determine if calcineurin promotes niche-to-mesenchymal cell state transitions to slow fin outgrowth. In parallel with thesis research, the applicant will pursue extensive scientific communication training and activities, mentoring and teaching experiences, professional development workshops and networking events, and a diversity-supporting leadership role in the Women in Graduate Science organization.

项目总结 该项目为申请者提供交叉细胞、发育和再生的博士培训 生物学。斑马鱼会再生切除的鳍，包括它们的骨骼，恢复到原来的大小。 无论受伤程度如何。然而，如何“知道”在达到正确大小后何时停止生长，这一点很难理解。 明白了。申请人的实验室最近提出了一种新的稳健鳍尺寸恢复模型，该模型基于 骨架几何定义的“生态位”种群的初始量及其通过细胞状态的逐渐耗尽 过渡。此外，研究小组还发现，经典的斑马鱼突变体长鳍过度生长是导致 通过Kcnh2a K+通道顺式异位表达。Long fint2中的Kcnh2a导致壁龛细胞耐受和 因此，在再生过程中过度生长。申请者的移植实验显示异位 间充质细胞谱系中的kcnh2a产生了足够的过度生长。然而，kcnh2a如何 破坏有序的生态位枯竭是未知的。已发表的研究表明，抑制钙依赖 磷酸酶钙调神经磷酸酶也会导致明显的鳍过度生长。申请人的新上位论实验 提示Kcnh2a在钙调神经磷酸酶的上游起作用。他们的新转基因钙离子报告识别动态钙离子 初步转录学显示的巢/间充质细胞水平取决于特定表达 钙离子通道。这导致了一个中心假设，即钙通道活性和动态钙水平激活 钙调神经磷酸酶信号转导促进巢到间充质细胞状态的转变和结束鳍的再生。这个 申请者将追求三个具体目标：1)确定Kcnh2a如何影响细胞内钙动态 鳍再生，2)确定影响再生鳍大小的钙离子通道，以及3)确定钙调神经磷酸酶 促进从壁龛到间充质细胞状态的转变，以减缓鳍的生长。在进行论文研究的同时， 申请者将进行广泛的科学交流培训和活动，指导和教学 经验、专业发展研讨会和网络活动，以及支持多样性的 女性在研究生科学组织中的领导作用。