1 Scientific RepoRts | 7: 1601 | DOI:10.1038/s41598-017-01519-4 www.nature.com/scientificreports An ovine hepatorenal fbrocystic model of a Meckel-like syndrome associated with dysmorphic primary cilia and TMEM67 mutations C. Stayner 1 , C. A. poole 2,6 , s. R. McGlashan 3 , M. pilanthananond 1 , R. Brauning 4 , D. Markie 1 , B. Lett 1 , L. Slobbe 1 , A. Chae 1 , A. C. Johnstone 5 , C. G. Jensen 3 , J. C. McEwan 4 , K. Dittmer 5 , K. parker 2 , A. Wiles 1 , W. Blackburne 1 , A. Leichter 1 , M. Leask 1 , A. pinnapureddy 1 , M. Jennings 2 , J. A. Horsfeld 1 , R. J. Walker 2 & M. R. Eccles 1 Meckel syndrome (MKS) is an inherited autosomal recessive hepatorenal fbrocystic syndrome, caused by mutations in TMEM67, characterized by occipital encephalocoele, renal cysts, hepatic fbrosis, 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 identifed in ovine TMEM67 were pathogenic in zebrafsh phenotype rescue assays. Meckelin protein was expressed in afected and unafected kidney epithelial cells by immunoblotting, and in primary cilia of lamb kidney cyst epithelial cells by immunofuorescence. In contrast to primary cilia of relatively consistent length and morphology in unafected kidney cells, those of afected 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 confrmed in cultured interstitial fbroblasts from afected kidneys. These primary cilia dysmorphologies and length control defects were signifcantly greater in afected cells compared to unafected controls. In conclusion, we describe abnormalities involving primary cilia length and morphology in the frst reported example of a large animal model of MKS, in which we have identifed TMEM67 mutations. Inherited renal cystic/fbrocystic diseases constitute an important subset of monogenic disorders, transmitted as autosomal dominant, autosomal recessive, or X-linked traits, and are responsible for more than 5% of world- wide end-stage renal disease 1 . Whereas the development of fuid-flled cysts and progressive impairment of renal function are common features, these disorders are distinguished from each other by diferent ages of onset, var- iable rates of renal disease progression, and a diverse array of extra-renal manifestations 1–3 . Te two major types of polycystic kidney disease (PKD) in humans have autosomal dominant (ADPKD) and autosomal recessive (ARPKD) inheritance 1, 4 . ADPKD is the most common dominant genetic disease in humans, afecting 1 in 500 individuals 1 and has a late onset. Recessive disorders include ARPKD 5 , nephronophthisis 6 , Meckel syndrome 7–9 , Joubert syndrome, Bardet-Biedl syndrome and other related disorders 10 . While less common than ADPKD, these 1 Department of Pathology, Dunedin School of Medicine, University of Otago, PO Box 56, Dunedin 9054, New Zealand. 2 Department of Medicine, Dunedin School of Medicine, University of Otago, PO Box 56, Dunedin 9054, New Zealand. 3 Department of Anatomy and Medical Imaging, The University of Auckland 1142, Private Bag, 92019, Auckland, New Zealand. 4 AgResearch Invermay Agricultural Centre, Mosgiel, 9053, New Zealand. 5 institute of Veterinary, Animal and Biomedical Sciences, Massey University, Tennant Drive, Palmerston North, 4472, New Zealand. 6 Present address: 150 Warren Street, Wanaka, 9305, New Zealand. C. Stayner and C. A. Poole contributed equally to this work. Correspondence and requests for materials should be addressed to M.R.E. (email: michael. eccles@otago.ac.nz) Received: 17 August 2016 Accepted: 29 March 2017 Published: xx xx xxxx opeN