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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要该项目为申请人提供了博士学位。训练与细胞，发育和再生相交生物学。斑马鱼再生切除的鳍，包括其骨射线骨架，回到原始尺寸无论受伤程度如何。然而，鳍如何“知道”何时停止随着正确的尺寸而停止生长很差理解。申请人的实验室最近提出了一种新的模型，以基于骨骼几何定义的“利基”种群的初始数量及其逐渐消耗的细胞状态过渡。此外，该小组在经典的斑马鱼突变体Longfint2中发现了巨大的鳍过度生长。通过KCNH2A K+通道的顺式分类表达。 longfint2中的kcnh2a导致利基细胞的渗透性和因此，再生期间超出产物。申请人的移植实验显示了生态生成小众细胞的间充质谱系中的KCNH2A足以过度生长。但是，如何kcnh2a 破坏有序的利基部署是未知的。已发表的研究表明抑制Ca2+依赖性磷酸酶钙调蛋白还会导致炎症的过度生长。申请人的新上毒实验建议钙调神经素上游的KCNH2A功能。他们的新转基因CA2+报告器确定了动态CA2+ 初步转录组学表明特定表达的小众/间质细胞中的水平 CA2+通道。这导致了一个中心假设，即Ca2+通道活性和动态Ca2+水平激活钙调神经蛋白信号传导促进生态界面至间质细胞状态转变和终止鳍片改革。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+动力学。鳍的再生，2）确定影响再生鳍大小的Ca2+通道，3）确定钙调神经酶是否是否促进壁层到间充质细胞状态的趋势，以减慢鳍片的生长。与论文研究并行申请人将进行广泛的科学沟通培训和活动，心理和教学经验，专业发展研讨会和网络活动以及多样性支持研究生科学组织中的女性领导角色。