Forkhead Factor, FOXL2, in Pituitary Development

描述(申请人提供)：foxL-2在发育中的眼睛和成年卵巢中表达，是人类这些器官正常发育和功能所必需的。最近，FOXL2在发育中的小鼠脑垂体中被检测到从e10.5到el5的表达。为了研究FOXL2在垂体发育中的作用，我们将进行以下研究：鉴定FOXL2的发育表达并将其置于垂体发育调控的遗传层次中；在细胞培养和转基因小鼠中确定FOXL2表达的重要调控序列；通过功能等位基因的获得和丧失来检测FOXL2在垂体前叶发育中的作用。将使用正常小鼠脑垂体的原位杂交来表征FOXL2的表达。FOXL2的正常表达模式将与关键垂体转录因子突变的小鼠进行比较。这一信息将把FOXL2放在遗传层次结构中。哺乳动物细胞的瞬时转基因和凝胶迁移率改变分析将被用来定义FOXL2表达的最小基本序列，并测试遗传层次中FOXL2上游因素的直接调控。这些发现将在转基因动物身上得到证实。为了描述FOXL2在垂体发育中的作用，FOXL2将在小鼠的垂体中过度表达并被特异性干扰，从而使我们能够将FOXI.2在垂体中的作用与其在其他器官中的作用分开。

DESCRIPTION (provided by applicant): Foxl-2 is expressed in the developing eye and in the adult ovary and is required for normal development and function of these organs in humans. Recently, expression of FoxL2 has been detected in the developing mouse pituitary from e10.5 to el 5. To examine the role of FOXL2 in pituitary development, the following studies will be performed: characterize the developmental expression of Foxl2 and place it in the genetic hierarchy of pituitary developmental control, define important regulatory sequences for Foxl2 expression in cell culture and transgenic mice, and examine the role of FOXL2 in the developing anterior pituitary by generation of gain and loss of function alleles. In situ hybridization of pituitaries from normal mice will be used to characterize Foxl2 expression. Normal expression patterns for Foxl2 will be compared to those in mice with mutations in critical pituitary transcription factors. This information will place FOXL2 in the genetic hierarchy. Transient transfections of mammalian cells and electrophoretic mobility shift assays will be used to define the minimal essential sequence for Foxl2 expression and to test for direct regulation by factors that are upstream of Foxl2 in the genetic hierarchy. These findings will be confirmed in transgenic animals. To delineate the role of FOXL2 in pituitary development, FoxL2 will be over-expressed and will be disrupted specifically in the pituitaries of mice, allowing us to separate the role of FOXI.2 in the pituitary from its role in other organs.