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

Research output: Contribution to journalArticleResearchpeer-review

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.

LanguageEnglish
Article numberbio036475
Pages1-7
Number of pages7
JournalBiology Open
Volume7
Issue number10
DOIs
Publication statusPublished - Oct 2018

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Neurodegenerative diseases
neurodegenerative diseases
Zebrafish
Danio rerio
Cell culture
Neurodegenerative Diseases
Neurons
cell culture
Cell Culture Techniques
neurons
Machado-Joseph Disease
genetically modified organisms
sarcoma
Agglomeration
Cytology
Sarcoma
Proteins
proteins
Amyotrophic Lateral Sclerosis
Pathology

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

Cite this

Acosta, Jamie R. ; Watchon, Maxinne ; Yuan, Kristy C. ; Fifita, Jennifer A. ; Svahn, Adam J. ; Don, Emily K. ; Winnick, Claire G. ; Blair, Ian P. ; Nicholson, Garth A. ; Cole, Nicholas J. ; Goldsbury, Claire ; Laird, Angela S. / Neuronal cell culture from transgenic zebrafish models of neurodegenerative disease. In: Biology Open. 2018 ; Vol. 7, No. 10. pp. 1-7.
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Neuronal cell culture from transgenic zebrafish models of neurodegenerative disease. / Acosta, Jamie R.; Watchon, Maxinne; Yuan, Kristy C.; Fifita, Jennifer A.; Svahn, Adam J.; Don, Emily K.; Winnick, Claire G.; Blair, Ian P.; Nicholson, Garth A.; Cole, Nicholas J.; Goldsbury, Claire; Laird, Angela S.

In: Biology Open, Vol. 7, No. 10, bio036475, 10.2018, p. 1-7.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Neuronal cell culture from transgenic zebrafish models of neurodegenerative disease

AU - Acosta, Jamie R.

AU - Watchon, Maxinne

AU - Yuan, Kristy C.

AU - Fifita, Jennifer A.

AU - Svahn, Adam J.

AU - Don, Emily K.

AU - Winnick, Claire G.

AU - Blair, Ian P.

AU - Nicholson, Garth A.

AU - Cole, Nicholas J.

AU - Goldsbury, Claire

AU - Laird, Angela S.

N1 - 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.

PY - 2018/10

Y1 - 2018/10

N2 - 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.

AB - 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.

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KW - Ataxin-3 (ATXN3)

KW - Fused in sarcoma (FUS)

KW - Primary neuronal cell culture

KW - Spinocerebellar ataxia type-3

KW - Transgenic zebrafish

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UR - http://purl.org/au-research/grants/nhmrc/1069235

UR - http://purl.org/au-research/grants/nhmrc/1146750

U2 - 10.1242/bio.036475

DO - 10.1242/bio.036475

M3 - Article

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