PUBLICATION

Neuromuscular regulation in zebrafish by a large AAA+ ATPase/ubiquitin ligase, mysterin/RNF213

Authors

Kotani, Y., Morito, D., Yamazaki, S., Ogino, K., Kawakami, K., Takashima, S., Hirata, H., Nagata, K.

ID

ZDB-PUB-151105-6

Date

2015

Source

Scientific Reports 5: 16161 (Journal)

Registered Authors

Hirata, Hiromi, Kawakami, Koichi

Keywords

Developmental biology, Molecular biology

MeSH Terms

Zebrafish/growth & development
Zebrafish/metabolism*
Zebrafish Proteins/antagonists & inhibitors
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Neovascularization, Physiologic
Animals, Genetically Modified/growth & development
Animals, Genetically Modified/metabolism
Humans
Muscle Fibers, Skeletal/metabolism
Muscle Fibers, Skeletal/pathology
Animals
Motor Neurons/metabolism
Oligonucleotides, Antisense/metabolism
Motor Activity
Ubiquitin-Protein Ligases/antagonists & inhibitors
Ubiquitin-Protein Ligases/genetics
Ubiquitin-Protein Ligases/metabolism*
RING Finger Domains
Microscopy, Electron
Larva/metabolism
Immunohistochemistry

(all 22)

PubMed

26530008 Full text @ Sci. Rep.

Abstract

Mysterin (also known as RNF213) is a huge intracellular protein with two AAA+ ATPase modules and a RING finger ubiquitin ligase domain. Mysterin was originally isolated as a significant risk factor for the cryptogenic cerebrovascular disorder moyamoya disease, and was found to be involved in physiological angiogenesis in zebrafish. However, the function and the physiological significance of mysterin in other than blood vessels remain largely unknown, although mysterin is ubiquitously expressed in animal tissues. In this study, we performed antisense-mediated suppression of a mysterin orthologue in zebrafish larvae and revealed that mysterin-deficient larvae showed significant reduction in fast myofibrils and immature projection of primary motoneurons, leading to severe motor deficits. Fast muscle-specific restoration of mysterin expression cancelled these phenotypes, and interestingly both AAA+ ATPase and ubiquitin ligase activities of mysterin were indispensable for proper fast muscle formation, demonstrating an essential role of mysterin and its enzymatic activities in the neuromuscular regulation in zebrafish.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
RFP	nkgSA2AzGFF598AGt; nkuasrfp1aTg	control	Long-pec	fast muscle cell	IHC	Fig. 4
RFP	nkgSA2AzGFF598AGt; nkuasrfp1aTg	control	Long-pec	fast muscle cell	IHC	Fig. 5
RFP	nkgSA2AzGFF598AGt; nkuasrfp1aTg + MO2-rnf213a	control	Long-pec	fast muscle cell	IHC	Fig. 5
RFP	nkgSA2AzGFF598AGt; nkuasrfp1aTg + MO2-rnf213a	control	Long-pec	fast muscle cell	IHC	Fig. 4
rnf213a	WT	control	Long-pec	whole organism	RTPCR	Fig. 1
rnf213a	WT + MO2-rnf213a	standard conditions	Long-pec	whole organism	RTPCR	Fig. 1

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + MO2-rnf213a	standard conditions	Prim-5	muscle pioneer skeletal muscle cell differentiation process quality, abnormal	Fig. 5
WT + MO2-rnf213a	standard conditions	Prim-5	myotome has extra parts of type muscle pioneer, abnormal	Fig. 5
WT + MO2-rnf213a	standard conditions	Prim-5	myotome has normal numbers of parts of type slow muscle cell, normal	Fig. 5
WT + MO2-rnf213a	standard conditions	Long-pec	CaP motoneuron axon extension involved in axon guidance decreased process quality, abnormal	Fig. 2
WT + MO2-rnf213a	standard conditions	Long-pec	fast muscle cell has fewer parts of type fast muscle cell skeletal muscle myofibril, abnormal	Fig. 3
WT + MO2-rnf213a	standard conditions	Long-pec	fast muscle cell skeletal muscle fiber development decreased process quality, abnormal	Fig. 3
WT + MO2-rnf213a	standard conditions	Long-pec	fast muscle cell Z disc increased thickness, abnormal	Fig. 3
WT + MO2-rnf213a	standard conditions	Long-pec	MiP motor neuron axon extension involved in axon guidance decreased process quality, abnormal	Fig. 2
WT + MO2-rnf213a	standard conditions	Long-pec	muscle pioneer skeletal muscle cell differentiation process quality, abnormal	Fig. 5
WT + MO2-rnf213a	standard conditions	Long-pec	myotome fast muscle cell loose, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Construct	Type
nkgSA2AzGFF598AGt	Gt(gSA2Az-GAL4FF)	Transgenic Insertion
nkuasrfp1aTg	Tg(5xUAS:RFP)	Transgenic Insertion
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion

1 - 3 of 3

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
rnf213a	MO1-rnf213a	MRPHLNO
rnf213a	MO2-rnf213a	MRPHLNO

Fish

Fish
nkgSA2AzGFF598AGt; nkuasrfp1aTg
nkgSA2AzGFF598AGt; nkuasrfp1aTg + MO2-rnf213a
WT
WT + MO2-rnf213a

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
RFP	EFG	RFP

Mapping

Kotani et al., 2015 ZDB-PUB-151105-6 PMID:26530008

Neuromuscular regulation in zebrafish by a large AAA+ ATPase/ubiquitin ligase, mysterin/RNF213

Kotani et al., 2015

ZDB-PUB-151105-6
PMID:26530008