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.

项目概要 该项目为申请人提供博士学位。训练交叉细胞、发育和再生 生物学。斑马鱼将切除的鳍（包括其骨射线骨骼）再生回原来的大小 与受伤程度无关。然而，当达到正确的尺寸时，鱼鳍如何“知道”何时停止生长却知之甚少。 明白了。申请人的实验室最近提出了一种基于稳健鳍尺寸恢复的新模型 骨骼几何定义的“利基”群体的初始量及其通过细胞状态的逐渐耗尽 过渡。此外，研究小组发现经典斑马鱼突变体 longfint2 中鳍的急剧过度生长是导致 通过 kcnh2a K+ 通道的顺式异位表达。 longfint2 中的 Kcnh2a 导致生态位细胞持久性和 因此在再生过程中过度生长。申请人的移植实验显示异位 产生微环境细胞的间充质谱系中的kcnh2a足以过度生长。然而，kcnh2a如何 破坏有序的生态位消耗尚不清楚。已发表的研究表明抑制 Ca2+ 依赖性 磷酸酶钙调神经磷酸酶也会导致鳍明显过度生长。申请人的新上位实验 表明 Kcnh2a 在钙调神经磷酸酶上游发挥作用。他们的新转基因 Ca2+ 报告基因识别动态 Ca2+ 初步转录组学表明的生态位/间充质细胞中的水平取决于具体表达 Ca2+ 通道。这导致了中心假设：Ca2+ 通道活动和动态 Ca2+ 水平激活 钙调神经磷酸酶信号传导促进生态位到间充质细胞状态转变和端鳍再生。这 申请人将追求三个具体目标：1) 确定 Kcnh2a 如何影响细胞内 Ca2+ 动态 鳍再生，2) 识别影响再生鳍尺寸的 Ca2+ 通道，以及 3) 确定钙调神经磷酸酶是否 促进生态位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， 申请人将进行广泛的科学传播培训和活动、指导和教学 经验、专业发展研讨会和社交活动，以及多元化支持 在研究生科学研究女性组织中发挥领导作用。