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.
| Language | English |
|---|---|
| Article number | bio036475 |
| Pages | 1-7 |
| Number of pages | 7 |
| Journal | Biology Open |
| Volume | 7 |
| Issue number | 10 |
| DOIs | |
| Publication status | Published - Oct 2018 |
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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
}
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 journal › Article › Research › peer-review
TY - JOUR
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.
KW - Amyotrophic lateral sclerosis (ALS)
KW - Ataxin-3 (ATXN3)
KW - Fused in sarcoma (FUS)
KW - Primary neuronal cell culture
KW - Spinocerebellar ataxia type-3
KW - Transgenic zebrafish
UR - http://www.scopus.com/inward/record.url?scp=85055194864&partnerID=8YFLogxK
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
VL - 7
SP - 1
EP - 7
JO - Biology Open
T2 - Biology Open
JF - Biology Open
SN - 2046-6390
IS - 10
M1 - bio036475
ER -