Roles of the kinase NUAK1 on spliceosome in the mouse cortex

The project focuses on the protein kinase NUAK1, which controls the development of brain cortical connectivity in mice. Recent data suggest that NUAK1 is involved in the regulation of the spliceosome and more broadly of mRNA biology. The aim of this project is to test whether inactivation of NUAK1 (constitutive KO) affects the expression level of genes relevant for cortical development in the embryonic brain. We collected the cortex of embryos (littermates) at the late embryonic developmental stage (E18.5, i.e. just before birth). The samples will be either KO or WT (controls). We collected the samples on the same day, under the same conditions, and as far as possible we made homogeneous groups: same litter, same proportion of males/females. We collected the cortex of embryos from a constitutive NUAK1 KO line, at the E18.5 embryonic stage. At this stage the cortex is mainly composed of maturing neurons (70X), migrating interneurons (10X) and progenitors/glial cells/endothelial cells (15-20X). Collection conditions: Samples were obtained from microdissected embryos. The females were mated at the same time and the embryos were from 4 litters.本项目聚焦于蛋白激酶NUAK1，该激酶可调控小鼠大脑皮层连接的发育进程。近期研究数据显示，NUAK1参与剪接体（spliceosome）的调控，并广泛涉及mRNA生物学过程。本项目旨在探究NUAK1组成型敲除（constitutive KO）是否会影响胚胎大脑中与皮层发育相关的基因表达水平。我们于胚胎发育晚期（即出生前，E18.5）收集了同窝胚胎的大脑皮层组织，样本分为敲除组（KO）与野生型对照组（WT）。所有样本均于同日、同一实验条件下采集，并尽可能构建同质性分组：采用同窝胚胎，且雌雄比例保持一致。我们从NUAK1组成型敲除小鼠品系中，于E18.5胚胎阶段收集了胚胎大脑皮层组织。该发育阶段的大脑皮层主要由三类细胞构成：成熟神经元（占比70%）、迁移中的中间神经元（占比10%）以及祖细胞/胶质细胞/内皮细胞（占比15%-20%）。样本收集条件：样本取自显微解剖后的胚胎；所有雌鼠均于同期交配，胚胎来源于4窝幼崽。