Restoration of renal function in zebrafish models of ciliopathies
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BRIEF REPORT
Restoration of renal function in zebrafish models of ciliopathies Jonathan L. Tobin & Philip L. Beales
Received: 21 February 2008 / Revised: 23 April 2008 / Accepted: 16 May 2008 / Published online: 5 July 2008 # The Author(s) 2008
Abstract The ciliopathies are a class of rare human genetic disease whose aetioligies lie in defective primary cilia. Typical ciliopathies include Bardet–Biedl syndrome (BBS), nephronophthisis (NPHP), Jeune, Joubert, oro-facial-digital (OFD1) and Meckel (MKS) syndromes. All ciliopathies have the common denominator of renal disease, often including tubular cysts. In this study, we have modelled a range of ciliopathies in zebrafish and shown in all cases that knocking down these genes causes cystic lesions in the kidney. We have identified two drugs, rapamycin and roscovitine, which ameliorate the renal phenotype, both morphologically and functionally. This is the first study in which zebrafish has been used to identify potential therapeutic modalities for ciliopathic renal disease, and the results pave the way for further investigations in mammalian models.
several ciliopathy genes (Bbs1, 2, 4, 6; Ofd1; Alms1, Pkhd), and they develop renal cysts but often not until several months of age [3]. The zebrafish is developing as a valid model for investigating human cystic kidney diseases [4]. The zebrafish pronephros develops after 2 days post-fertilisation (dpf), and the presence of cysts is easily screened at 3 dpf. These cysts often compromise renal function, resulting in body oedema by 4 dpf. Transparency of the embryos makes morphological scoring rapid, and the large clutches enables scalability. Ex vivo development allows therapeutic compounds to be added directly to the water in which the eggs are incubated [5]. Our aim is to create fish models for most of the known human ciliopathies, screen them for a kidney phenotype and then try to identify compounds that could prevent cyst formation and potentially restore function.
Keywords Ciliopathy . Kidney cysts . Rapamycin . Roscovitine . Therapeutics
Methods
Introduction Primary cilia function to detect signalling molecules and mechanical stimuli, such as fluid flow. Mutations in genes encoding some of the approximately 1000 ciliary proteins cause ciliopathies. Renal disease is a common denominator of ciliopathies, often presenting with tubular cysts, lobulation and clubbing [1, 2]. Published mouse models exist for J. L. Tobin : P. L. Beales (*) Molecular Medicine Unit, UCL Institute of Child Health, 30 Guilford Street, London WC1N 1EH, UK e-mail: [email protected]
Between 2 and 4.5 ng of translation and splice-blocking morpholino oligonucleotides (MOs) (Gene Tools, Philomath, OR) were used against the following genes (disease caused by human mutations in brackets): bbs4, bbs6, bbs8 (Bardet– Biedl syndrome); nphp2 (nephronophthisis); nphp5 (Senior– Loken syndrome); nphp6 (Joubert syndrome); mks1, mks3 (Meckel syndrome); ofd1 (oro-facial-digital syndrome type I). The MO sequences are available on request. Renal lesion sizes
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