Gene expression and protein distribution of ADAMTSL-4 in human iris, choroid and retina
Gene expression and protein distribution of ADAMTSL-4 in human iris, choroid and retina
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DOI：
10.1136/bjophthalmol-2013-303353
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发表时间：
2013-09-01
影响因子：
4.1
通讯作者：
Charteris, David G.
中科院分区：
文献类型：
--
作者：
Chandra, Aman;Jones, Megan;Charteris, David G.
背景 ADAMTSL4 突变最近被证明是常染色体隐性遗传孤立性晶状体异位 (IEL) 的主要原因。然而，该蛋白质的功能和眼部定位尚未完全确定。因此，我们旨在证实该基因和蛋白在正常眼组织中的表达。方法在死后48小时内获取供体眼组织，并分离虹膜、脉络膜和视网膜进行分析。使用逆转录 PCR 检查四只眼睛中编码 ADAMTSL4 的 mRNA 的表达。还通过蛋白质印迹分析在两只眼睛中研究了该分子的蛋白质编码。此外，在对该蛋白进行免疫染色并对染色组织进行共聚焦分析后，在整个眼睛的低温切片中研究了 ADAMTSL4 的原位定位。 结果 ADAMTSL4 的 mRNA 和编码蛋白均被证明在虹膜和脉络膜组织中表达，但在神经视网膜中不表达。共聚焦研究表明ADAMTS-Like 4存在于睫状体和睫状突以及视网膜色素上皮中。结论我们已经证实了ADAMTSL4在人眼组织中的基因和蛋白表达。表达模式可能表明该基因的进一步功能超出了其在 IEL 中的致病作用所暗示的功能。
Background 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.Methods Donor ocular tissue was obtained within 48h 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.Results 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.Conclusions 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.