CONTROL OF CORNEAL ENDOTHELIUM DEVELOPMENT IN THE MOUSE

小鼠角膜内皮发育的控制

• 批准号: 6395008
• 负责人: BRIGID L.M. HOGAN
• 金额: $ 3.2万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6395008
• 关键词: Mus musculus; biological signal transduction; cadherins; cell differentiation; cell migration; corneal endothelium; electron microscopy; electrophysiology; embryo /fetus tissue /cell culture; embryogenesis; epidermal growth factor; gene expression; growth factor receptors; histogenesis; immunocytochemistry; in situ hybridization; lens; mesenchyme; mutant; neural crest; polymerase chain reaction; protein structure function; subtraction hybridization; transcription factor; transforming growth factors

The goal of this study is to define the role of Mf1, a forehead/winged helix transcription factor, in the development of the cornea of the mammalian eye. We have previously shown that in mouse embryos lacking a functional Mf1 gene (generated either by homologous recombination-Mf1/lacZ-or as a result of spontaneous mutation-Mf1/Ch) the corneal endothelium fails to differentiate from the embryonic corneal mesenchyme. This is accompanied by a number of other eve developmental abnormalities, including failure of anterior chamber formation, iris dystrophy, and abnormalities of the trabecular meshwork (Kidson et al, 1999, Appendix). The failure to form a corneal endothelium in these mutants thus provides an ideal system for investigate the normal development of the corneal endothelium in vivo, about which little is known. The human homologue of the mouse Mf1 gene (known as FKHL/FREAC3) has recently been cloned by two groups. Dominant mutations in this gene have been shown to be associated with inherited anterior segment defects, including hypoplasia of the iris, dysgenesis of the anterior angle and trabecular meshwork, corneal opacity and increased risk of juvenile onset congenital glaucoma (Axenfeld-Rieger Anomaly, Mears et al, 1998: Nishimura et al, 1998). In addition, several inherited disorders of the cornea have been reported to humans. For example, congenital hereditary endothelial dystrophy, a condition that can be either autosomal dominant or recessive, is associated with absence of the endothelium, thickening of the stroma and corneal opacity (Mullany et al, 1995: Kirkness et al, 1987). Thus, the proposed studies will lead to a better understanding of human eye defects associated with corneal endothelium failure, including glaucoma and corneal endothelial dystrophy. The hypothesis will be tested that Mf1 functions in the corneal mesenchyme as part of the downstream signaling pathway from as yet unidentified factors secreted by the embryonic lens. The techniques to be used will include in vitro culture and differentiation of normal and mutant corneal mesenchyme, co-culture with embryonic lens, addition of growth factors, and screening for Mf1 regulated genes.

这项研究的目的是定义MF1（额头/有翼螺旋转录因子）在哺乳动物眼角膜发展中的作用。我们先前已经表明，在缺乏功能性MF1基因的小鼠胚胎中（由于自发突变-MF1/CH而由同源重组MF1/LACZ-OR生成）角膜内皮无法与胚胎角膜间质分化。这伴随着许多其他夏娃发育异常，包括前腔形成，虹膜营养不良症和小梁网术的异常（Kidson等，1999，附录）。因此，在这些突变体中未能形成角膜内皮，因此为研究体内角膜内皮的正常发育提供了理想的系统，这一点鲜为人知。小鼠MF1基因的人类同源物（称为FKHL/FREAC3）最近被两组克隆。 Dominant mutations in this gene have been shown to be associated with inherited anterior segment defects, including hypoplasia of the iris, dysgenesis of the anterior angle and trabecular meshwork, corneal opacity and increased risk of juvenile onset congenital glaucoma (Axenfeld-Rieger Anomaly, Mears et al, 1998: Nishimura et al, 1998).此外，已经向人类报告了几种角膜的遗传疾病。例如，先天性遗传性内皮营养不良症（这种可能是常染色体显性效果或隐性的疾病）与缺乏内皮，基质的增厚和角膜不透明度有关（Mullany等，1995：Kirkness等，1987）。因此，提出的研究将使人们对与角膜内皮衰竭相关的人眼缺陷有更好的了解，包括青光眼和角膜内皮营养不良。该假设将测试，即MF1在角膜间充质中起作用，这是下游信号通路的一部分，从胚胎透镜分泌的AS尚不确定的因素。要使用的技术将包括正常和突变角膜间充质和突变的角膜间充质，与胚胎晶状体共培养的体外培养和分化，加入生长因子以及MF1调节基因的筛选。