Expression of ALS/FTD-linked mutant CCNF in zebrafish leads to increased cell death in the spinal cord and an aberrant motor phenotype

Alison L. Hogan, Emily K. Don, Stephanie L. Rayner, Albert Lee, Angela S. Laird, Maxinne Watchon, Claire Winnick, Ingrid S. Tarr, Marco Morsch, Jennifer A Fifita, Serene S. L. Gwee, Isabel Formella, Elinor Hortle, Kristy C. Yuan, Mark P. Molloy, Kelly L. Williams, Garth A. Nicholson, Roger S. Chung, Ian P. Blair, Nicholas J. Cole

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, fatal neurodegenerative disease characterised by the death of upper and lower motor neurons. Approximately 10% of cases have a known family history of ALS and disease-linked mutations in multiple genes have been identified. ALS-linked mutations in CCNF were recently reported, however the pathogenic mechanisms associated with these mutations are yet to be established. To investigate possible disease mechanisms, we developed in vitro and in vivo models based on an ALS-linked missense mutation in CCNF. Proteomic analysis of the in vitro models identified the disruption of several cellular pathways in the mutant model, including caspase-3 mediated cell death. Transient overexpression of human CCNF in zebrafish embryos supported this finding, with fish expressing the mutant protein found to have increased levels of cleaved (activated) caspase-3 and increased cell death in the spinal cord. The mutant CCNF fish also developed a motor neuron axonopathy consisting of shortened primary motor axons and increased frequency of aberrant axonal branching. Importantly, we demonstrated a significant correlation between the severity of the CCNF-induced axonopathy and a reduced motor response to a light stimulus (photomotor response). This is the first report of an ALS-linked CCNF mutation in vivo and taken together with the in vitro model identifies the disruption of cell death pathways as a significant consequence of this mutation. Additionally, this study presents a valuable new tool for use in ongoing studies investigating the pathobiology of ALS-linked CCNF mutations.

LanguageEnglish
Pages2616-2626
Number of pages11
JournalHuman molecular genetics
Volume26
Issue number14
Early online date21 Apr 2017
DOIs
Publication statusPublished - 15 Jul 2017

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Amyotrophic Lateral Sclerosis
Zebrafish
Spinal Cord
Cell Death
Phenotype
Mutation
Motor Neurons
Caspase 3
Fishes
Missense Mutation
Mutant Proteins
Neurodegenerative Diseases
Proteomics
Axons
Embryonic Structures
Light
Genes
In Vitro Techniques

Bibliographical note

A corrigendum exists for this artice and can be found in Human Molecular Genetics (2019), Volume 28(4).Page 698 at doi: 10.1093/hmg/ddy362

Cite this

@article{de72a09064904d45851a028f718f4295,
title = "Expression of ALS/FTD-linked mutant CCNF in zebrafish leads to increased cell death in the spinal cord and an aberrant motor phenotype",
abstract = "Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, fatal neurodegenerative disease characterised by the death of upper and lower motor neurons. Approximately 10{\%} of cases have a known family history of ALS and disease-linked mutations in multiple genes have been identified. ALS-linked mutations in CCNF were recently reported, however the pathogenic mechanisms associated with these mutations are yet to be established. To investigate possible disease mechanisms, we developed in vitro and in vivo models based on an ALS-linked missense mutation in CCNF. Proteomic analysis of the in vitro models identified the disruption of several cellular pathways in the mutant model, including caspase-3 mediated cell death. Transient overexpression of human CCNF in zebrafish embryos supported this finding, with fish expressing the mutant protein found to have increased levels of cleaved (activated) caspase-3 and increased cell death in the spinal cord. The mutant CCNF fish also developed a motor neuron axonopathy consisting of shortened primary motor axons and increased frequency of aberrant axonal branching. Importantly, we demonstrated a significant correlation between the severity of the CCNF-induced axonopathy and a reduced motor response to a light stimulus (photomotor response). This is the first report of an ALS-linked CCNF mutation in vivo and taken together with the in vitro model identifies the disruption of cell death pathways as a significant consequence of this mutation. Additionally, this study presents a valuable new tool for use in ongoing studies investigating the pathobiology of ALS-linked CCNF mutations.",
author = "Hogan, {Alison L.} and Don, {Emily K.} and Rayner, {Stephanie L.} and Albert Lee and Laird, {Angela S.} and Maxinne Watchon and Claire Winnick and Tarr, {Ingrid S.} and Marco Morsch and Fifita, {Jennifer A} and Gwee, {Serene S. L.} and Isabel Formella and Elinor Hortle and Yuan, {Kristy C.} and Molloy, {Mark P.} and Williams, {Kelly L.} and Nicholson, {Garth A.} and Chung, {Roger S.} and Blair, {Ian P.} and Cole, {Nicholas J.}",
note = "A corrigendum exists for this artice and can be found in Human Molecular Genetics (2019), Volume 28(4).Page 698 at doi: 10.1093/hmg/ddy362",
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Expression of ALS/FTD-linked mutant CCNF in zebrafish leads to increased cell death in the spinal cord and an aberrant motor phenotype. / Hogan, Alison L.; Don, Emily K.; Rayner, Stephanie L.; Lee, Albert; Laird, Angela S.; Watchon, Maxinne; Winnick, Claire; Tarr, Ingrid S.; Morsch, Marco; Fifita, Jennifer A; Gwee, Serene S. L.; Formella, Isabel; Hortle, Elinor; Yuan, Kristy C.; Molloy, Mark P.; Williams, Kelly L.; Nicholson, Garth A.; Chung, Roger S.; Blair, Ian P.; Cole, Nicholas J.

In: Human molecular genetics, Vol. 26, No. 14, 15.07.2017, p. 2616-2626.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Expression of ALS/FTD-linked mutant CCNF in zebrafish leads to increased cell death in the spinal cord and an aberrant motor phenotype

AU - Hogan, Alison L.

AU - Don, Emily K.

AU - Rayner, Stephanie L.

AU - Lee, Albert

AU - Laird, Angela S.

AU - Watchon, Maxinne

AU - Winnick, Claire

AU - Tarr, Ingrid S.

AU - Morsch, Marco

AU - Fifita, Jennifer A

AU - Gwee, Serene S. L.

AU - Formella, Isabel

AU - Hortle, Elinor

AU - Yuan, Kristy C.

AU - Molloy, Mark P.

AU - Williams, Kelly L.

AU - Nicholson, Garth A.

AU - Chung, Roger S.

AU - Blair, Ian P.

AU - Cole, Nicholas J.

N1 - A corrigendum exists for this artice and can be found in Human Molecular Genetics (2019), Volume 28(4).Page 698 at doi: 10.1093/hmg/ddy362

PY - 2017/7/15

Y1 - 2017/7/15

N2 - Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, fatal neurodegenerative disease characterised by the death of upper and lower motor neurons. Approximately 10% of cases have a known family history of ALS and disease-linked mutations in multiple genes have been identified. ALS-linked mutations in CCNF were recently reported, however the pathogenic mechanisms associated with these mutations are yet to be established. To investigate possible disease mechanisms, we developed in vitro and in vivo models based on an ALS-linked missense mutation in CCNF. Proteomic analysis of the in vitro models identified the disruption of several cellular pathways in the mutant model, including caspase-3 mediated cell death. Transient overexpression of human CCNF in zebrafish embryos supported this finding, with fish expressing the mutant protein found to have increased levels of cleaved (activated) caspase-3 and increased cell death in the spinal cord. The mutant CCNF fish also developed a motor neuron axonopathy consisting of shortened primary motor axons and increased frequency of aberrant axonal branching. Importantly, we demonstrated a significant correlation between the severity of the CCNF-induced axonopathy and a reduced motor response to a light stimulus (photomotor response). This is the first report of an ALS-linked CCNF mutation in vivo and taken together with the in vitro model identifies the disruption of cell death pathways as a significant consequence of this mutation. Additionally, this study presents a valuable new tool for use in ongoing studies investigating the pathobiology of ALS-linked CCNF mutations.

AB - Amyotrophic lateral sclerosis (ALS) is a rapidly progressive, fatal neurodegenerative disease characterised by the death of upper and lower motor neurons. Approximately 10% of cases have a known family history of ALS and disease-linked mutations in multiple genes have been identified. ALS-linked mutations in CCNF were recently reported, however the pathogenic mechanisms associated with these mutations are yet to be established. To investigate possible disease mechanisms, we developed in vitro and in vivo models based on an ALS-linked missense mutation in CCNF. Proteomic analysis of the in vitro models identified the disruption of several cellular pathways in the mutant model, including caspase-3 mediated cell death. Transient overexpression of human CCNF in zebrafish embryos supported this finding, with fish expressing the mutant protein found to have increased levels of cleaved (activated) caspase-3 and increased cell death in the spinal cord. The mutant CCNF fish also developed a motor neuron axonopathy consisting of shortened primary motor axons and increased frequency of aberrant axonal branching. Importantly, we demonstrated a significant correlation between the severity of the CCNF-induced axonopathy and a reduced motor response to a light stimulus (photomotor response). This is the first report of an ALS-linked CCNF mutation in vivo and taken together with the in vitro model identifies the disruption of cell death pathways as a significant consequence of this mutation. Additionally, this study presents a valuable new tool for use in ongoing studies investigating the pathobiology of ALS-linked CCNF mutations.

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U2 - 10.1093/hmg/ddx136

DO - 10.1093/hmg/ddx136

M3 - Article

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EP - 2626

JO - Human molecular genetics

T2 - Human molecular genetics

JF - Human molecular genetics

SN - 0964-6906

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