RNA Seq analysis of GCNF mutant and wt in anterior and posterior cranial regions of E9.25 embryos in Mus musculus

Neural crest cells are migratory progenitor cells that contribute to nearly all tissues and organs throughout the body. Their formation, migration and differentiation are regulated by a multitude of signaling pathways, that when disrupted can lead to disorders termed neurocristopathies. While work in avian and amphibian species has revealed essential factors governing the specification and induction of neural crest cells during gastrulation and neurulation in non-mammalian species, their functions do not appear to be conserved in mice, leaving major gaps in our understanding of neural crest cell formation in mammals. Here we describe Germ Cell Nuclear Factor (GCNF/Nr6a1), an orphan nuclear receptor, as a critical regulator of neural crest cell formation in mice. Gcnf null mutant mice, exhibit a major disruption of neural crest cell formation. The purpose of this experiment is to examine gene expression changes in response to Gcnf mutation in anterior and posterior cranial regions of E9.25 mouse embryos. Mouse embryos were collected at E9.25 corresponding to 9-11 somite stage separating the cranial regions (1st pharyngeal arch and anterior) and posterior regions (2nd pharyngeal arch and posterior) for comparison of gene expression between homozygous GCNF mutant and wt.神经嵴细胞（Neural crest cells）是一类迁移性祖细胞，可参与体内几乎所有组织与器官的形成。其形成、迁移与分化受多种信号通路调控；当这些通路功能异常时，会引发一类被称为神经嵴病（neurocristopathies）的疾病。尽管在鸟类与两栖类动物中的研究已揭示了非哺乳类物种原肠胚形成（gastrulation）与神经胚形成（neurulation）过程中，调控神经嵴细胞特化与诱导的关键因子，但此类因子的功能在小鼠中并不保守，这使得我们对哺乳类神经嵴细胞形成机制的认知存在重大空白。本研究将生殖细胞核因子（Germ Cell Nuclear Factor，GCNF/Nr6a1）——一种孤儿核受体（orphan nuclear receptor）——鉴定为小鼠神经嵴细胞形成的关键调控因子。Gcnf纯合缺失突变小鼠表现出神经嵴细胞形成的严重缺陷。本实验旨在检测E9.25小鼠胚胎颅前区与颅后区内，Gcnf突变所诱导的基因表达变化。研究收集了对应9-11体节期（somite stage）的E9.25小鼠胚胎，将其颅区划分为颅前区（第一鳃弓pharyngeal arch及前部区域）与颅后区（第二鳃弓及后部区域），以比较纯合GCNF突变体与野生型（wild type，wt）样本间的基因表达差异。