mtor Haploinsufficiency Ameliorates Renal Cysts and Cilia Abnormality in Adult Zebrafish tmem67 Mutants
- 11 February 2021
- journal article
- research article
- Published by Ovid Technologies (Wolters Kluwer Health) in Journal of the American Society of Nephrology
- Vol. 32 (4), 822-836
- https://doi.org/10.1681/asn.2020070991
Abstract
Background Although zebrafish embryos have been used to study ciliogenesis and model polycystic kidney disease (PKD), adult zebrafish remain unexplored. Methods Transcription activator-like effector nucleases (TALEN) technology was used to generate mutant for tmem67, the homolog of the mammalian causative gene for Meckel syndrome type 3 (MKS3). Classic 2D and optical-clearing 3D imaging of an isolated adult zebrafish kidney were used to examine cystic and ciliary phenotypes. A hypomorphic mtor strain or rapamycin was used to inhibit mTOR activity. Results Adult tmem67 zebrafish developed progressive mesonephric cysts that share conserved features of mammalian cystogenesis, including a switch of cyst origin with age and an increase in proliferation of cyst-lining epithelial cells. The mutants had shorter and fewer distal single cilia and greater numbers of multiciliated cells (MCCs). Absence of a single cilium preceded cystogenesis, and expansion of MCCs occurred after pronephric cyst formation and was inversely correlated with the severity of renal cysts in young adult zebrafish, suggesting a primary defect and an adaptive action, respectively. Finally, the mutants exhibited hyperactive mTOR signaling. mTOR inhibition ameliorated renal cysts in both the embryonic and adult zebrafish models; however, it only rescued ciliary abnormalities in the adult mutants. Conclusions Adult zebrafish tmem67 mutants offer a new vertebrate model for renal cystic diseases, in which cilia morphology can be analyzed at a single-nephron resolution and mTOR inhibition proves to be a candidate therapeutic strategy.Keywords
Funding Information
- Mayo Clinic (NIDDK DK90728)
- Mayo Foundation for Medical Education and Research
- Mayo Clinic Center for Biomedical Discovery
This publication has 80 references indexed in Scilit:
- A transition zone complex regulates mammalian ciliogenesis and ciliary membrane compositionNature Genetics, 2011
- Ciliogenesis: building the cell's antennaNature Reviews Molecular Cell Biology, 2011
- Identification of adult nephron progenitors capable of kidney regeneration in zebrafishNature, 2011
- Craniofacial ciliopathies: A new classification for craniofacial disordersAmerican Journal of Medical Genetics Part A, 2010
- NovelTMEM67mutations and genotype-phenotype correlates in meckelin-related ciliopathiesHuman Mutation, 2010
- Identification of Signaling Pathways Regulating Primary Cilium Length and Flow-Mediated AdaptationCurrent Biology, 2010
- Impaired Wnt–β-catenin signaling disrupts adult renal homeostasis and leads to cystic kidney ciliopathyNature Medicine, 2009
- Autosomal dominant polycystic kidney disease: the last 3 yearsKidney International, 2009
- Mutation spectrum of Meckel syndrome genes: one group of syndromes or several distinct groups?Human Mutation, 2009
- Intraflagellar transportNature Reviews Molecular Cell Biology, 2002