ORBIT, UCL Institute of Ophthalmology, London, UK.
Br J Ophthalmol. 2013 Sep;97(9):1208-12. doi: 10.1136/bjophthalmol-2013-303353. Epub 2013 Jul 11.
Mutations in ADAMTSL4 have recently been shown to be the major cause of autosomal recessive isolated ectopia lentis (IEL). However, the function and ocular localisation of the protein is yet to be fully established. We therefore aimed to confirm the expression of this gene and protein in normal ocular tissue.
Donor ocular tissue was obtained within 48 h post-mortem and iris, choroid and retina were isolated for analysis. Expression of mRNA coding for ADAMTSL4 was examined in four eyes using reverse transcription PCR. Protein coding for this molecule was also investigated in two eyes by western blot analysis. Furthermore, the in situ localisation of ADAMTSL4 was investigated in cryostat sections of whole eyes following immunostaining for this protein and confocal analysis of the stained tissue.
mRNA and protein coding for ADAMTSL4 were both demonstrated to be expressed in iris and choroidal tissue but were absent from the neural retina. Confocal studies revealed ADAMTS-Like 4 to be present in the ciliary body and ciliary processes and also in the retinal pigment epithelium.
We have confirmed the gene and protein expression of ADAMTSL4 in human ocular tissue. The pattern of expression may suggest further functions of this gene beyond those suggested by its causative role in IEL.
最近的研究表明,ADAMTSL4 基因突变是常染色体隐性孤立性晶状体异位(IEL)的主要原因。然而,该蛋白的功能和眼部定位尚未完全确定。因此,我们旨在确认该基因和蛋白在正常眼部组织中的表达。
在死后 48 小时内获取供体眼部组织,并分离虹膜、脉络膜和视网膜进行分析。使用逆转录 PCR 在四只眼中检查 ADAMTSL4 编码 mRNA 的表达。通过 Western blot 分析在两只眼中研究该分子的蛋白质编码。此外,通过对该蛋白进行免疫染色并对染色组织进行共聚焦分析,研究 ADAMTSL4 在整个眼部冷冻切片中的原位定位。
mRNA 和蛋白质编码 ADAMTSL4 在虹膜和脉络膜组织中均有表达,但在神经视网膜中不存在。共聚焦研究显示 ADAMTS-Like 4 存在于睫状体和睫状突以及视网膜色素上皮中。
我们已经证实 ADAMTSL4 在人眼部组织中的基因和蛋白表达。这种表达模式可能表明该基因的功能超出了其在 IEL 中的致病作用所暗示的功能。