PUBLICATION
Neuronal cell culture from transgenic zebrafish models of neurodegenerative disease
- Authors
- Acosta, J.R., Watchon, M., Yuan, K.C., Fifita, J., Svahn, A.J., Don, E.K., Blair, I.P., Nicholson, G.A., Cole, N.J., Goldsbury, C., Laird, A.S.
- ID
- ZDB-PUB-180908-1
- Date
- 2018
- Source
- Biology Open 7(10): (Journal)
- Registered Authors
- Cole, Nicholas, Don, Emily, Laird, Angela, Svahn, Adam, Watchon, Maxinne, Yuan, Kristy
- Keywords
- Amyotrophic Lateral Sclerosis (ALS), Ataxin-3 (ATXN3), Fused in Sarcoma (FUS), Primary neuronal cell culture, Spinocerebellar ataxia type-3, Transgenic zebrafish
- MeSH Terms
- none
- PubMed
- 30190267 Full text @ Biol. Open
Citation
Acosta, J.R., Watchon, M., Yuan, K.C., Fifita, J., Svahn, A.J., Don, E.K., Blair, I.P., Nicholson, G.A., Cole, N.J., Goldsbury, C., Laird, A.S. (2018) Neuronal cell culture from transgenic zebrafish models of neurodegenerative disease. Biology Open. 7(10):.
Abstract
We describe a protocol for culturing neurons from transgenic zebrafish embryos to investigate the subcellular distribution and protein aggregation status of neurodegenerative disease-causing proteins. The utility of the protocol was demonstrated on cell cultures from zebrafish that transgenically express disease-causing variants, human FUS and ataxin-3 proteins, in order to study amyotrophic lateral sclerosis (ALS) and spinocerebellar ataxia type-3 (SCA3), respectively. A mixture of neuronal subtypes, including motor neurons, exhibited differentiation and neurite outgrowth in the cultures. As reported previously, mutant human FUS was found to be mislocalized from nuclei to the cytosol, mimicking the pathology seen in human ALS and the zebrafish FUS model. In contrast, neurons cultured from zebrafish expressing human ataxin-3 with disease-associated expanded polyQ repeats did not accumulate within nuclei in a manner often reported to occur in SCA3. Despite this, the subcellular localisation of human ataxin-3 protein seen in the cell cultures was similar to that found in the SCA3 zebrafish themselves. The finding of similar protein localisation and aggregation status in the neuronal cultures and corresponding transgenic zebrafish models confirms that this cell culture model is a useful tool for investigating the cell biology and proteinopathy signatures of mutant proteins for the study of neurodegenerative disease.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping