An ovine hepatorenal fibrocystic model of a Meckel-like syndrome associated with dysmorphic primary cilia and TMEM67 mutations.
An ovine hepatorenal fibrocystic model of a Meckel-like syndrome associated with dysmorphic primary cilia and TMEM67 mutations.
复制标题
DOI：
10.1038/s41598-017-01519-4
复制
发表时间：
2017-05-09
影响因子：
4.6
通讯作者：
Eccles MR
中科院分区：
文献类型：
--
作者：
Stayner C;Poole CA;McGlashan SR;Pilanthananond M;Brauning R;Markie D;Lett B;Slobbe L;Chae A;Johnstone AC;Jensen CG;McEwan JC;Dittmer K;Parker K;Wiles A;Blackburne W;Leichter A;Leask M;Pinnapureddy A;Jennings M;Horsfield JA;Walker RJ;Eccles MR
Meckel综合征（MKS）是一种遗传性常染色体隐性肝肾纤维囊性综合征，由TMEM 67突变引起，以枕部脑膨出、肾囊肿、肝纤维化和多指（趾）畸形为特征。在这里，我们描述了一个绵羊模型MKS，肾脏和肝脏异常，没有多指或枕脑腔突出。纯合子错义p.（Ile 681 Asn; Ile 687 Ser）在绵羊TMEM 67中鉴定的突变在斑马鱼表型拯救试验中是致病性的。免疫印迹法检测到Meckelin蛋白在受累和未受累的肾上皮细胞中表达，免疫荧光法检测到Meckelin蛋白在羔羊肾囊肿上皮细胞原代纤毛中表达。与未受影响的肾细胞中相对一致的长度和形态的初级纤毛相反，受影响的囊肿衬里细胞显示出一系列短和极长的纤毛，以及异常的形态，如沿着轴丝的球根状区域。假定的纤毛片段也始终位于囊肿腔内容物内。异常纤毛表型在受影响肾脏的培养间质成纤维细胞中得到进一步证实。与未受影响的对照组相比，受影响细胞的这些初级纤毛畸形和长度控制缺陷显着更大。总之，我们描述的异常涉及初级纤毛长度和形态的第一个报告的例子，一个大型动物模型的MKS，其中我们已经确定了TMEM 67突变。
Meckel syndrome (MKS) is an inherited autosomal recessive hepatorenal fibrocystic syndrome, caused by mutations in TMEM67, characterized by occipital encephalocoele, renal cysts, hepatic fibrosis, and polydactyly. Here we describe an ovine model of MKS, with kidney and liver abnormalities, without polydactyly or occipital encephalocoele. Homozygous missense p.(Ile681Asn; Ile687Ser) mutations identified in ovine TMEM67 were pathogenic in zebrafish phenotype rescue assays. Meckelin protein was expressed in affected and unaffected kidney epithelial cells by immunoblotting, and in primary cilia of lamb kidney cyst epithelial cells by immunofluorescence. In contrast to primary cilia of relatively consistent length and morphology in unaffected kidney cells, those of affected cyst-lining cells displayed a range of short and extremely long cilia, as well as abnormal morphologies, such as bulbous regions along the axoneme. Putative cilia fragments were also consistently located within the cyst luminal contents. The abnormal ciliary phenotype was further confirmed in cultured interstitial fibroblasts from affected kidneys. These primary cilia dysmorphologies and length control defects were significantly greater in affected cells compared to unaffected controls. In conclusion, we describe abnormalities involving primary cilia length and morphology in the first reported example of a large animal model of MKS, in which we have identified TMEM67 mutations.