Distinguishing the role of ERC1 isoforms in membrane trafficking during craniofacial and neuronal development

项目摘要/摘要 膜转运是导致细胞形状改变和组织完整性的重要细胞机制 在开发过程中。损害正常运输的突变会导致发育障碍，影响 头面部形态和神经功能。临床报告确认了颅面部 与12号染色体末端缺失相关的表型和神经发育障碍。ERc1(ELKS， Rab6IP2，CAST)，是一种膜转运的调节因子，在 删除了12号染色体区域。然而，ERC1在调节颅面和神经元中的作用 发展并不容易确定，特别是由于多个可选择剪接的ERC1亚型。 据报道，人类erc1亚型有不同的表达，因此，可能有不同的细胞 功能。与其他脊椎动物模型不同，斑马鱼有两个平行的erc1基因，erc1a和erc1b，两者都是 它们与选择性剪接的人类ERC1亚型同源。通过正向遗传学筛查或 在斑马鱼中进行CRISPR/Cas9基因组编辑，我们建议研究erc1同源基因在头面部中的作用 发育过程中的软骨和脑的形态。这个项目的目的是a)识别斑马鱼erc1 Paralog在发育过程中的头面部软骨功能和神经组织形态，以及b)确定 Erc1类似物通过Rab GTP酶相互作用在细胞形态和细胞存活中的作用 发展。该项目使用活体脊椎动物建模和成像技术来绘制不同的 人类ERC1亚型对颅面和神经元发育的功能贡献。调查 不同ERC1亚型在细胞形态和发育过程中存活的分子功能将提供 颅面畸形与神经发育障碍共病的细胞学基础 发生在12号染色体远端缺失的患者中。

Project Summary/Abstract Membrane trafficking is an essential cellular mechanism contributing to cell shape changes and tissue integrity during development. Mutations impairing proper trafficking can result in developmental disorders, affecting both craniofacial morphology and neurologic function. Clinical reports have identified both craniofacial phenotypes and neurodevelopmental disorders associated with distal chromosome 12 deletions. ERC1 (ELKS, Rab6IP2, CAST), a regulator of membrane trafficking, is found within the smallest region of overlap between deleted chromosome 12 regions. However, a role for ERC1 in regulating craniofacial and neuronal development has not been easy to determine, particularly due to multiple alternatively spliced ERC1 isoforms. Human ERC1 isoforms are reported to be differentially expressed, and therefore, could have distinct cellular function. Unlike other vertebrate models, zebrafish have two paralogous erc1 genes, erc1a and erc1b, both of which are homologous to alternatively spliced human ERC1 isoforms. By forward genetics screen or CRISPR/Cas9 genome editing in zebrafish, we propose to examine the role of erc1 paralogs in craniofacial cartilage and brain morphology during development. The aims of this project are to a) identify zebrafish erc1 paralogs’ function in craniofacial cartilage and neural tissue morphology during development, and b) determine Erc1 paralogs’ function in establishing cell shape and cell survival through Rab GTPase interaction during development. This project uses in vivo vertebrate animal modeling and imaging techniques to map distinct functional contribution of human ERC1 isoforms to craniofacial and neuronal development. Investigating the molecular function of separate ERC1 isoforms in cell shape and survival during development will provide insight into a cellular basis to craniofacial dysmorphology and neurodevelopmental disorder comorbidity occurring in patients with distal chromosome 12 deletions.