Combining Zebrafish and Mouse Models to Test the Function of Deubiquitinating Enzyme (Dubs) Genes in Development: Role of USP45 in the Retina.
Ubiquitination is a dynamic and reversible posttranslational modification. Much effort has been devoted to characterize the function of ubiquitin pathway genes in the cell context, but much less is known on their functional role in the development and maintenance of organs and tissues in the organism. In fact, several ubiquitin ligases and deubiquitinating enzymes (DUBs) are implicated in human pathological disorders, from cancer to neurodegeneration. The aim of our work is to explore the relevance of DUBs in retinal function in health and disease, particularly since some genes related to the ubiquitin or SUMO pathways cause retinal dystrophies, a group of rare diseases that affect 1:3000 individuals worldwide. We propose zebrafish as an extremely useful and informative genetic model to characterize the function of any particular gene in the retina, and thus complement the expression data from mouse. A preliminary characterization of gene expression in mouse retinas (RT-PCR and in situ hybridization) was performed to select particularly interesting genes, and we later replicated the experiments in zebrafish. As a proof of concept, we selected ups45 to be knocked down by morpholino injection in zebrafish embryos. Morphant phenotypic analysis showed moderate to severe eye morphological defects, with a defective formation of the retinal structures, therefore supporting the relevance of DUBs in the formation and differentiation of the vertebrate retina, and suggesting that genes encoding ubiquitin pathway enzymes are good candidates for causing hereditary retinal dystrophies.
Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, Avda. Diagonal 643, 08028, Barcelona, Spain.
Vasileios
V
Toulis
Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, Avda. Diagonal 643, 08028, Barcelona, Spain.|Department of Human Genetics, and Donders Center for Neurosciences, Radboud University Medical Centre, Nijmegen, The Netherlands.
Alejandro
A
Garanto
Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, Avda. Diagonal 643, 08028, Barcelona, Spain. gmarfany@ub.edu.|Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III, Barcelona, Spain. gmarfany@ub.edu.|Institut de Biomedicina de la Universitat de Barcelona (IBUB), Barcelona, Spain. gmarfany@ub.edu.
Gemma
G
Marfany
D000818:Animals; D000072017:Deubiquitinating Enzymes / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000201:enzymology / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology; D017403:In Situ Hybridization; D051379:Mice; D012160:Retina / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism
D029961:Zebrafish Proteins; D044767:Ubiquitin-Protein Ligases; D000072017:Deubiquitinating Enzymes
Deubiquitinating enzymes; Morpholino knockdown; Neurodegeneration; Retina; USP45; Zebrafish Animal Model