Microarray Analysis of GCNF mutant and wt E9.0 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 E9.0 mouse embryos. Homozygous wt and mutant GCNF mouse embryos at developmental stage E9.0 were isolated in duplicate for RNA extraction and hybridization on Affymetrix microarrays.神经嵴细胞（Neural crest cells）是一类具有迁移能力的祖细胞，参与机体几乎所有组织与器官的形成。其形成、迁移及分化受多条信号通路调控，当这些通路发生异常时，可引发一类被称为神经嵴病（neurocristopathies）的疾病。尽管在非哺乳动物类群的鸟类与两栖动物研究中，已有成果揭示了原肠胚形成与神经管闭合阶段调控神经嵴细胞特化与诱导的关键因子，但这些因子的功能在小鼠中似乎并不保守，这使得我们对哺乳动物神经嵴细胞形成机制的认知存在重大空白。本研究将孤核受体生殖细胞核因子（Germ Cell Nuclear Factor, GCNF/Nr6a1）鉴定为小鼠神经嵴细胞形成的关键调控因子。Gcnf基因敲除突变小鼠会出现神经嵴细胞形成的严重缺陷。本实验旨在探究发育阶段为E9.0的小鼠胚胎中，Gcnf突变所诱导的基因表达变化。我们将发育至E9.0阶段的纯合野生型（wild type, wt）与纯合Gcnf突变型小鼠胚胎各重复取样两份，用于RNA提取及Affymetrix基因芯片杂交实验。