Zhou G, Price C E, Rosenquist T H, Gadson P F, Godfrey M
Munroe Center for Human Genetics, University of Nebraska Medical Center, Omaha 68198-5430, USA.
In Vitro Cell Dev Biol Anim. 2000 Jan;36(1):19-25. doi: 10.1290/1071-2690(2000)036<0019:pcasoc>2.0.co;2.
The recent identification of numerous matrix genes and gene products has allowed a detailed examination of their roles in development. Two of these extracellular matrix proteins, fibrillin-1 and fibrillin-2, are components of the elastin-associated microfibrils. Given what is known about the distribution of the fibrillins in normal tissues and the abnormalities that result when mutations occur, a basic hypothesis has emerged: fibrillin-1 is primarily responsible for load bearing and providing structural integrity, whereas fibrillin-2 may be a director of elastogenesis. Nevertheless, examination of phenotypes in disorders caused by mutations in fibrillin-1 or fibrillin-2 suggests some common functions. To better understand these similar and diverse roles, it would be helpful to examine these proteins during chick development. To accomplish this goal, it is first necessary to characterize the chick homologs of the known fibrillins. In this study, the partial chick FBN1 cDNA was identified by polymerase chain reaction-aided cloning as a first step toward elucidating these goals. Sequence analysis indicated that there is striking conservation between chick and mammalian fibrillin-1 at the DNA and protein levels. Antisense and sense riboprobes were synthesized and used in in situ hybridization in stage 14 chick embryos and high levels of FBN1 transcripts were observed in the heart.
最近大量基质基因和基因产物的鉴定使得人们能够详细研究它们在发育过程中的作用。其中两种细胞外基质蛋白,原纤蛋白-1和原纤蛋白-2,是与弹性蛋白相关的微原纤维的组成成分。鉴于已知原纤蛋白在正常组织中的分布情况以及突变发生时所导致的异常情况,一个基本假设应运而生:原纤蛋白-1主要负责承受负荷并提供结构完整性,而原纤蛋白-2可能是弹性生成的主导者。然而,对由原纤蛋白-1或原纤蛋白-2突变引起的疾病中的表型进行研究表明它们具有一些共同功能。为了更好地理解这些相似和不同的作用,在鸡的发育过程中研究这些蛋白质将有所帮助。为了实现这一目标,首先有必要鉴定已知原纤蛋白的鸡同源物。在本研究中,通过聚合酶链反应辅助克隆鉴定了鸡FBN1 cDNA的部分序列,这是朝着阐明这些目标迈出的第一步。序列分析表明,鸡和哺乳动物的原纤蛋白-1在DNA和蛋白质水平上存在显著的保守性。合成了反义核糖探针和正义核糖探针,并将其用于14期鸡胚的原位杂交,结果在心脏中观察到了高水平的FBN1转录本。