Intracellular pH Dynamics in Zebrafish Cranial Neural Crest Development

斑马鱼颅神经嵴发育的细胞内 pH 动态

基本信息

批准号：
10604726
负责人：
Cambria Chou-Freed
金额：
$ 4.36万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-05 至 2025-09-04
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10604726
关键词：
Address Behavior Biology Cell Adhesion Cell Fate Control Cell membrane Cells Cephalic Chick Chondrocytes Craniofacial Abnormalities Data Defect Development Disease Drosophila genus Embryo Embryonic Development Exhibits Focal Adhesion Kinase 1 Future Genetic Genetic Transcription Health Human Image Intestines Ions Lead Modeling Molecular Mus Neural Crest Neural Crest Cell Odontoblasts Organism Osteocytes Ovarian Follicle Paraxial Mesoderm Pharmacology Population Proteins Regulation Reporter Reporting Role Specific qualifier value System Talin Testing Time Tissues Zebrafish base blastomere structure cell behavior cell fate specification cell motility cell type cofilin craniofacial craniofacial development embryonic stem cell epithelial to mesenchymal transition in vivo innovation insight interest lens live cell imaging migration non-genetic novel protonation ratiometric sensor single-cell RNA sequencing stem cells tissue repair

项目摘要

PROJECT SUMMARY/ABSTRACT Intracellular pH (pHi) dynamics are a critical regulator of cell fate changes. Our lab and others demonstrated that an increase in pHi is necessary for the differentiation and lineage specification of multiple cell types, including mouse embryonic stem cells, Drosophila ovarian follicle stem cells, mouse intestinal stem cells, and chick paraxial mesoderm. Our lab has also shown how pHi dynamics regulate cell adhesion and migration behaviors by modifying the protonation state of pH-sensing proteins, including -catenin, cofilin, talin, and focal adhesion kinase. However, the mechanisms underlying pHi regulation of cell fate changes remain unresolved. Furthermore, our understanding of pHi dynamics during embryonic development in vivo is limited, largely due to lack of appropriate models. To address this, I generated a novel system for interrogating the functional significance of pHi dynamics during embryonic development in vivo using zebrafish, a genetically tractable organism suited for in vivo live cell imaging of embryos. Using this new model, I will determine the role of pHi dynamics in the development of cranial neural crest (NC), a highly conserved vertebrate embryonic cell population that gives rise to diverse cell types, including chondrocytes, osteocytes, and odontoblasts. Many cell behaviors involved in cranial NC development are regulated by pHi dynamics in other cell types, including lineage specification, cell migration, and epithelial to mesenchymal transition. Thus, cranial NC represents an ideal model for addressing the gaps in our understanding of pHi dynamics during embryonic development in vivo. By following early NC cells through the differentiation of cranial lineages, I will test the central hypothesis that pHi dynamics regulate cranial NC development, at the stage of delamination, migration, or lineage specification. In Aim 1, I will resolve spatial and temporal pHi dynamics during zebrafish cranial NC development by in vivo live cell imaging. My preliminary data indicate a higher pHi in migratory compared with premigratory cranial NC cells. In Aim 2, I will experimentally perturb pHi in zebrafish NC cells through pharmacologic and genetic modulation of plasma membrane ion transporters and determine the effect on cranial NC cell behaviors and transcription, thus establishing the functional significance of pHi dynamics during zebrafish cranial NC development. My findings have promise to reveal new insight on the cellular and molecular factors controlling craniofacial development, with important implications for human congenital diseases and tissue repair.

项目概要/摘要我们的实验室和其他实验室证明，细胞内 pH (pHi) 动态是细胞命运变化的关键调节因素。 pHi 的增加对于多种细胞类型的分化和谱系规范是必要的，包括小鼠胚胎干细胞、果蝇卵泡干细胞、小鼠肠道干细胞和我们的实验室还展示了 pHi 动力学如何调节细胞粘附和迁移。通过改变 pH 传感蛋白（包括 -catenin、cofilin、talin 和focal）的质子化状态来改变行为然而，pHi 调节细胞命运变化的机制仍未解决。此外，我们对体内胚胎发育过程中 pHi 动力学的理解是有限的，这很大程度上是由于为了解决这个问题，我生成了一个新的系统来询问功能。使用斑马鱼（一种遗传上易于处理的动物）体内胚胎发育过程中 pHi 动态的重要性适合胚胎活体细胞成像的生物体使用这个新模型，我将确定 pHi 的作用。高度保守的脊椎动物胚胎细胞颅神经嵴（NC）发育动态产生多种细胞类型的群体，包括软骨细胞、骨细胞和成牙本质细胞。参与颅脑 NC 发育的细胞行为受其他细胞类型的 pHi 动力学调节，包括因此，颅内 NC 代表了谱系规范、细胞迁移和上皮间质转化。解决我们对胚胎发育过程中 pHi 动态的理解差距的理想模型通过追踪早期 NC 细胞的颅骨谱系分化，我将测试中枢。假设 pHi 动力学在分层阶段调节颅脑 NC 发育，在目标 1 中，我将解决迁移期间的空间和时间 pHi 动力学问题。通过体内活细胞成像进行斑马鱼颅脑 NC 发育我的初步数据表明 pHi 较高。与迁移前的颅内 NC 细胞相比，在目标 2 中，我将通过实验扰乱斑马鱼的 pHi。 NC细胞通过质膜离子转运蛋白的药理和遗传调节并确定对颅内 NC 细胞行为和转录的影响，从而确定 pHi 的功能意义我的研究结果有望揭示斑马鱼颅脑 NC 发育过程中的动态。控制颅面发育的细胞和分子因素，对人类具有重要意义先天性疾病和组织修复。