Neuronal cell culture from transgenic zebrafish models of neurodegenerative disease

Jamie R. Acosta, Maxinne Watchon, Kristy C. Yuan, Jennifer A. Fifita, Adam J. Svahn, Emily K. Don, Claire G. Winnick, Ian P. Blair, Garth A. Nicholson, Nicholas J. Cole, Claire Goldsbury, Angela S. Laird*

*Corresponding author for this work

Research output: Contribution to journalArticle

2 Citations (Scopus)
14 Downloads (Pure)

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 of human fused in sarcoma (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 localization of the human ataxin-3 protein seen in cell cultures was similar to that found in the SCA3 zebrafish themselves. The finding of similar protein localization 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.

Original languageEnglish
Article numberbio036475
Pages (from-to)1-7
Number of pages7
JournalBiology Open
Volume7
Issue number10
DOIs
Publication statusPublished - Oct 2018

Bibliographical note

Copyright 2018. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • Amyotrophic lateral sclerosis (ALS)
  • Ataxin-3 (ATXN3)
  • Fused in sarcoma (FUS)
  • Primary neuronal cell culture
  • Spinocerebellar ataxia type-3
  • Transgenic zebrafish

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