PUBLICATION

Zebrafish as a Model to Investigate Dynamin 2-Related Diseases

Authors

Bragato, C., Gaudenzi, G., Blasevich, F., Pavesi, G., Maggi, L., Giunta, M., Cotelli, F., Mora, M.

ID

ZDB-PUB-160205-3

Date

2016

Source

Scientific Reports 6: 20466 (Journal)

Registered Authors

Cotelli, Franco

Keywords

Genetic testing, Neuromuscular disease

MeSH Terms

Charcot-Marie-Tooth Disease/genetics
Charcot-Marie-Tooth Disease/pathology*
Dynamin II/deficiency*
Dynamin II/genetics*
Dynamin II/metabolism
Gene Knockdown Techniques
Muscle Cells/metabolism
Muscle Cells/pathology
Disease Models, Animal*
Sequence Homology, Nucleic Acid
Animals, Genetically Modified
Animals
Zebrafish/genetics
Zebrafish/metabolism*
Myopathies, Structural, Congenital/genetics
Myopathies, Structural, Congenital/pathology*
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism
Dynamins/genetics*
Dynamins/metabolism
Alternative Splicing
Humans
Mutation

(all 23)

PubMed

26842864 Full text @ Sci. Rep.

Abstract

Mutations in the dynamin-2 gene (DNM2) cause autosomal dominant centronuclear myopathy (CNM) and dominant intermediate Charcot-Marie-Tooth (CMT) neuropathy type B (CMTDIB). As the relation between these DNM2-related diseases is poorly understood, we used zebrafish to investigate the effects of two different DNM2 mutations. First we identified a new alternatively spliced zebrafish dynamin-2a mRNA (dnm2a-v2) with greater similarity to human DNM2 than the deposited sequence. Then we knocked-down the zebrafish dnm2a, producing defects in muscle morphology. Finally, we expressed two mutated DNM2 mRNA by injecting zebrafish embryos with human mRNAs carrying the R522H mutation, causing CNM, or the G537C mutation, causing CMT. Defects arose especially in secondary motor neuron formation, with incorrect branching in embryos injected with CNM-mutated mRNA, and total absence of branching in those injected with CMT-mutated mRNA. Muscle morphology in embryos injected with CMT-mutated mRNA appeared less regularly organized than in those injected with CNM-mutated mRNA. Our results showing, a continuum between CNM and CMTDIB phenotypes in zebrafish, similarly to the human conditions, confirm this animal model to be a powerful tool to investigate mutations of DNM2 in vivo.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
dnm2a	AB	standard conditions	14-19 somites	adaxial cell	ISH	Fig. 2
dnm2a	AB	standard conditions	14-19 somites	adaxial cell	ISH	Fig. S2
dnm2a	AB	standard conditions	10-13 somites	adaxial cell	ISH	Fig. S2
dnm2a	AB	standard conditions	20-25 somites	adaxial cell	ISH	Fig. S2
dnm2a	AB	standard conditions	5-9 somites	adaxial cell	ISH	Fig. S2
dnm2a	AB	standard conditions	14-19 somites	midbrain hindbrain boundary	ISH	Fig. 2
dnm2a	AB	standard conditions	Prim-5	midbrain hindbrain boundary	ISH	Fig. 2
dnm2a	AB	standard conditions	Prim-5	neural tube	ISH	Fig. 2
dnm2a	AB	standard conditions	14-19 somites	neural tube	ISH	Fig. 2
dnm2a	AB	standard conditions	14-19 somites	otic vesicle	ISH	Fig. 2

1 - 10 of 23

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + MO3-dnm2a	standard conditions	Protruding-mouth	skeletal muscle cell disorganized, abnormal	Fig. 4
AB + MO3-dnm2a	standard conditions	Protruding-mouth	somite hypoplastic, abnormal	Fig. 3
AB + MO3-dnm2a	standard conditions	Protruding-mouth	somitogenesis disrupted, abnormal	Fig. 3
AB + MO3-dnm2a	standard conditions	Protruding-mouth	startle response decreased process quality, abnormal	text only
AB + MO3-dnm2a	standard conditions	Protruding-mouth	thigmotaxis decreased process quality, abnormal	text only
AB + MO3-dnm2a	standard conditions	Day 4	skeletal muscle cell disorganized, abnormal	Fig. 3
AB + MO3-dnm2a	standard conditions	Day 4	skeletal muscle cell increased diameter, abnormal	Fig. 3
AB + MO3-dnm2a	standard conditions	Day 4	skeletal muscle cell morphology, abnormal	Fig. 3
AB + MO3-dnm2a	standard conditions	Day 4	skeletal muscle cell nucleus increased amount, abnormal	Fig. 3
AB + MO3-dnm2a	standard conditions	Day 4	skeletal muscle cell skeletal muscle myofibril disorganized, abnormal	Fig. 3

1 - 10 of 21

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Mutations / Transgenics

Human Disease / Model

Human Disease	Fish	Conditions	Evidence
centronuclear myopathy			TAS
Charcot-Marie-Tooth disease intermediate type			TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
dnm2a	MO3-dnm2a	MRPHLNO
dnm2a	MO4-dnm2a	MRPHLNO

Fish

Fish
AB
AB + MO3-dnm2a
AB + MO4-dnm2a

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Bragato et al., 2016 ZDB-PUB-160205-3 PMID:26842864

Zebrafish as a Model to Investigate Dynamin 2-Related Diseases

Bragato et al., 2016

ZDB-PUB-160205-3
PMID:26842864