Embryonic Development of the Cerebellum

描述（由申请人提供）：我们以前的工作集中在胚胎小脑颗粒细胞的规格。在这里，我们希望把注意力转向小脑的另一个主要神经元，浦肯野细胞和深层神经元。本项目的总体目标是确定四类转录因子LIM同源结构域基因的地形表达，（Lhx 1、Lhx 3、Lhx 5和Lhx 9），碱性螺旋-环-螺旋基因（Math 1，Mash 1，Ngn 1，Ngn 2），胚胎小脑原基中的浦肯野和深层神经元祖细胞群体中的含有配对盒的基因（Pax 2）和偶跳基因（Evx 1，2），并使用这些标记来检查影响细胞命运的因素。在鸡胚和Dreher突变小鼠的基因表达研究中，将测试局部诱导因子、骨形成蛋白对基因表达和细胞规范模式的作用。首先，我们将定义小鼠和禽类胚胎中的基因表达模式，重点关注深层神经元和浦肯野细胞产生的时间段。我们将通过研究另一个项目中产生的Lhx 1、Lhx 3、Lhx 5、Lhx 9、Math 1、Mash 1、Ngn 1、Ngn 2和Evx 1的BAC转基因小鼠来进一步完善我们的分析。其次，我们将研究Dreher突变小鼠，因为它遭受BMPs的损失，并在禽胚胎中使用卵内电穿孔来确定BMP 6，BMP 7和GDF 7在转录因子表达模式中的作用。最后，我们将研究这些转录因子在小脑细胞分化中的功能，通过分析小鼠靶向无效突变，并通过将吗啉代反义寡核苷酸导入鸟类胚胎。

DESCRIPTION (provided by applicant): Our previous work has focused on the specification of granule cells in the embryonic cerebellum. Here we wish to turn our attention to the other principal neuron of the cerebellum, the Purkinje cell and to the deep neurons. The overall aim of this project is to determine the topographic expression of four classes of transcription factors, LIM homeodomain genes (Lhx1, Lhx3, Lhx5, and Lhx9), basic helix-loop-helix genes (Math1, Mash1, Ngn1, Ngn2), Paired box containing genes (Pax2) and Even-Skipped genes (Evx1,2) in Purkinje and deep neuron progenitor cell populations in the embryonic cerebellar primordium, and to use these markers to examine factors that shape cell fate specification. The action of local inductive factors, BMPs on gene expression and cell specification patterning will be tested in gene expression studies in the chick embryo and in the Dreher mutant mouse.First, we will define patterns of gene expression in murine and avian embryos focusing on the time periods when deep neurons and Purkinje cells are generated. We will further refine our analysis by studying BAC transgenic mice for Lhx1, Lhx3, Lhx5, Lhx9, Math1, Mash1, Ngn1, Ngn2 and Evx1 generated in another project. Second, we will study the Dreher mutant mouse, because it suffers a loss of BMPs, and use in ovo electroporation in avian embryos to define the role of BMP6, BMP7 and GDF7 in transcription factor expression patterns. Finally, we will study the function of these transcription factors in cerebellar cell differentiation by analyzing mice with targeted null mutations and by transfecting morpholino antisense oligonucleotides into avian embryos.