Wani M A, Wert S E, Lingrel J B
Department of Molecular Genetics, Biochemistry and Microbiology College of Medicine, University of Cincinnati, Cincinnati, Ohio 45267-0524, USA.
J Biol Chem. 1999 Jul 23;274(30):21180-5. doi: 10.1074/jbc.274.30.21180.
Lung Kruppel-like factor (LKLF) is a member of the Kruppel-like factor family of transcription factors and is highly expressed in lung with limited distribution in other tissues. Mice lacking LKLF due to inactivation of LKLF by gene targeting die in utero at midgestation around day 12.5 due to severe hemorrhage, making it difficult to study the role of this transcription factor in lung development and function. However, in vitro organ culture of lung buds removed from 11.5-day-old LKLF(-/-) embryos show normal tracheobronchial tree formation. To examine later stages of lung development, the embryonic lethality due to germ line LKLF null mutation was circumvented by constructing LKLF homozygous null mouse embryonic stem cells, using a two-step gene targeting procedure, and determining whether these cells give rise to lung tissue. The targeted cells were used to produce chimeric animals, and the contribution of LKLF-deficient cells to the formation of various internal organs was analyzed. In chimeric mice that survived after birth, null embryonic stem cells contributed significantly to all of the major organs except the lungs. On the other hand, some highly chimeric animals died at birth, and histopathological examination of their lungs suggested abnormalities in their lung development. These studies show that LKLF plays an important role in normal lung development.
肺 Kruppel 样因子(LKLF)是 Kruppel 样转录因子家族的成员,在肺中高度表达,在其他组织中的分布有限。由于基因靶向导致 LKLF 失活而缺乏 LKLF 的小鼠在妊娠中期约 12.5 天死于子宫内,原因是严重出血,这使得研究这种转录因子在肺发育和功能中的作用变得困难。然而,从 11.5 天大的 LKLF(-/-)胚胎中取出的肺芽进行体外器官培养显示气管支气管树形成正常。为了研究肺发育的后期阶段,通过构建 LKLF 纯合无效小鼠胚胎干细胞,采用两步基因靶向程序,并确定这些细胞是否能产生肺组织,从而规避了由于种系 LKLF 无效突变导致的胚胎致死性。将靶向细胞用于产生嵌合动物,并分析 LKLF 缺陷细胞对各种内部器官形成的贡献。在出生后存活的嵌合小鼠中,无效胚胎干细胞对除肺以外的所有主要器官都有显著贡献。另一方面,一些高度嵌合的动物在出生时死亡,对其肺的组织病理学检查表明其肺发育异常。这些研究表明 LKLF 在正常肺发育中起重要作用。