PUBLICATION

CHCHD10^P80L knock-in zebrafish display a mild ALS-like phenotype

Authors

Légaré, V.P., Harji, Z.A., Rampal, C.J., Antonicka, H., Gurberg, T.J.N., Persia, O., Rodríguez, E.C., Shoubridge, E.A., Armstrong, G.A.B.

ID

ZDB-PUB-240912-7

Date

2024

Source

Experimental neurology 382: 114945 (Journal)

Registered Authors

Armstrong, Gary A.B.

Keywords

Als, CHCHD10, Knock-in mutation, Mitochondria, Motor neuron, Neurodegeneration, Zebrafish

MeSH Terms

Motor Neurons/metabolism
Motor Neurons/pathology
Neuromuscular Junction/genetics
Neuromuscular Junction/metabolism
Neuromuscular Junction/pathology
Phenotype*
Animals, Genetically Modified
Amyotrophic Lateral Sclerosis*/genetics
Amyotrophic Lateral Sclerosis*/pathology
Zebrafish*
Animals
Zebrafish Proteins/genetics
Disease Models, Animal
Mitochondrial Proteins*/genetics
Gene Knock-In Techniques

(all 15)

PubMed

39260590 Full text @ Exp. Neurol.

Abstract

Mutations in the nuclear-encoded mitochondrial gene CHCHD10 have been observed in patients with a spectrum of diseases that include amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). To investigate the pathogenic nature of disease-associated variants of CHCHD10 we generated a zebrafish knock-in (KI) model expressing the orthologous ALS-associated CHCHD10^P80L variant (zebrafish: Chchd10^P83L). Larval chchd10^P83L/P83L fish displayed reduced Chchd10 protein expression levels, motor impairment, reduced survival and abnormal neuromuscular junctions (NMJ). These deficits were not accompanied by changes in transcripts involved in the integrated stress response (ISR), phenocopying previous findings in our knockout (chchd10^-/-). Adult, 11-month old chchd10^P83L/P83L zebrafish, displayed smaller slow- and fast-twitch muscle cell cross-sectional areas compared to wild type zebrafish muscle cells. Motoneurons in the spinal cord of chchd10^P83L/P83L zebrafish displayed similar cross-sectional areas to that of wild type motor neurons and significantly fewer motor neurons were observed when compared to chchd2^-/- adult spinal cords. Bulk RNA sequencing using whole spinal cords of 7-month old fish revealed transcriptional changes associated with neuroinflammation, apoptosis, amino acid metabolism and mt-DNA inflammatory response in our chchd10^P83L/P83L model. The findings presented here, suggest that the CHCHD10^P80L variant confers an ALS-like phenotype when expressed in zebrafish.

Genes / Markers

Figures

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Expression

Gene	Antibody	Fish	Conditions	Stage	Anatomy	Assay	Figure
atf4a		chchd10^mg4/mg4 (TL)	standard conditions	Day 5	whole organism	RTPCR	Fig. 4
atf4a		TL	standard conditions	Day 5	whole organism	RTPCR	Fig. 4
atf4b		chchd10^mg4/mg4 (TL)	standard conditions	Day 5	whole organism	RTPCR	Fig. 4
atf4b		TL	standard conditions	Day 5	whole organism	RTPCR	Fig. 4
atf5a		chchd10^mg4/mg4 (TL)	standard conditions	Day 5	whole organism	RTPCR	Fig. 4
atf5a		TL	standard conditions	Day 5	whole organism	RTPCR	Fig. 4
atf5b		chchd10^mg4/mg4 (TL)	standard conditions	Day 5	whole organism	RTPCR	Fig. 4
atf5b		TL	standard conditions	Day 5	whole organism	RTPCR	Fig. 4
chchd10	Ab1-chchd10	chchd10^mg2/mg2 (TL)	standard conditions	Adult	brain	WB	Fig. 5
chchd10	Ab1-chchd10	chchd10^mg4/mg4 (TL)	standard conditions	Adult	brain	WB	Fig. 5

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
chchd10^mg2/mg2 (TL)	standard conditions	Adult	motor neuron mitochondrial crista morphology, normal	Fig. 7
chchd10^mg2/mg2 (TL)	standard conditions	Adult	spinal cord has normal numbers of parts of type motor neuron, normal	Fig. 7
chchd10^mg2/mg2 (TL)	standard conditions	Adult	swimming normal process quality, normal	Fig. 5
chchd10^mg2/mg2 (TL)	standard conditions	Adult	trunk decreased area, abnormal	Fig. 6
chchd10^mg4/mg4 (TL)	standard conditions	Long-pec	myotome neuromuscular junction development, skeletal muscle fiber decreased process quality, abnormal	Fig. 2
chchd10^mg4/mg4 (TL)	standard conditions	Long-pec	swimming decreased process quality, abnormal	Fig. 2
chchd10^mg4/mg4 (TL)	standard conditions	Long-pec	whole organism morphology, normal	Fig. 2
chchd10^mg4/mg4 (TL)	standard conditions	Day 5	swimming decreased process quality, abnormal	Fig. 3
chchd10^mg4/mg4 (TL)	standard conditions	Day 5	whole organism chchd10 expression decreased amount, abnormal	Fig. 1
chchd10^mg4/mg4 (TL)	standard conditions	Days 14-20 to Days 21-29	whole organism semi-viable, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Type	Affected Genomic Region
mg2	Insertion	chchd10
mg3	Small Deletion	chchd2
mg4	Complex	chchd10

Human Disease / Model

Human Disease	Fish	Conditions	Evidence
amyotrophic lateral sclerosis	chchd10^mg4/mg4 (TL)	standard conditions	TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
chchd10	CRISPR2-chchd10	CRISPR

Fish

Fish
chchd2^mg3/mg3 (TL)
chchd10^mg2/mg2 (TL)
chchd10^mg4/mg4 (TL)
TL

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-chat	polyclonal	chata		Goat
Ab1-chchd10	polyclonal	chchd10	IgG	Rabbit
Ab1-mt-co1	monoclonal		IgG2a	Mouse
Ab1-ndufa9	monoclonal		IgG	Mouse
AB1-sdhb	monoclonal	sdhb	IgG2a	Mouse
Ab1-uqcrc2	polyclonal		IgG	Rabbit
Ab1-vcl	monoclonal		IgG1	Mouse
Ab4-atp5a	monoclonal		IgG2b	Mouse

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Orthology

Engineered Foreign Genes

Mapping

Légaré et al., 2024 ZDB-PUB-240912-7 PMID:39260590

CHCHD10P80L knock-in zebrafish display a mild ALS-like phenotype

Légaré et al., 2024

ZDB-PUB-240912-7
PMID:39260590

CHCHD10^P80L knock-in zebrafish display a mild ALS-like phenotype