PUBLICATION

Myosin phosphatase Fine-tunes Zebrafish Motoneuron Position during Axonogenesis

Authors

Bremer, J., Granato, M.

ID

ZDB-PUB-161118-2

Date

2016

Source

PLoS Genetics 12: e1006440 (Journal)

Registered Authors

Granato, Michael

Keywords

Axons, Embryos, Myosins, Notochords, Spinal cord, Muscle cells, Motor neurons, Zebrafish

MeSH Terms

Muscle Cells/cytology
Muscle Cells/metabolism
Myosin-Light-Chain Phosphatase/biosynthesis
Myosin-Light-Chain Phosphatase/genetics*
Embryonic Development/genetics*
Notochord/growth & development
Zebrafish/genetics
Zebrafish/growth & development
Neurogenesis/genetics*
Animals
Motor Neurons/cytology
Motor Neurons/metabolism
Axons*
Spinal Cord/growth & development
Gene Expression Regulation, Developmental

(all 15)

PubMed

27855159 Full text @ PLoS Genet.

Abstract

During embryogenesis the spinal cord shifts position along the anterior-posterior axis relative to adjacent tissues. How motor neurons whose cell bodies are located in the spinal cord while their axons reside in adjacent tissues compensate for such tissue shift is not well understood. Using live cell imaging in zebrafish, we show that as motor axons exit from the spinal cord and extend through extracellular matrix produced by adjacent notochord cells, these cells shift several cell diameters caudally. Despite this pronounced shift, individual motoneuron cell bodies stay aligned with their extending axons. We find that this alignment requires myosin phosphatase activity within motoneurons, and that mutations in the myosin phosphatase subunit mypt1 increase myosin phosphorylation causing a displacement between motoneuron cell bodies and their axons. Thus, we demonstrate that spinal motoneurons fine-tune their position during axonogenesis and we identify the myosin II regulatory network as a key regulator.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
EGFP	nkuasgfp1aTg; p188Tg; smb1226Tg	standard conditions	20-25 somites to Prim-15	interneuron	IFL	Fig. 1
EGFP	nkuasgfp1aTg; p188Tg; smb1226Tg	standard conditions	20-25 somites to Prim-15	notochord	IFL	Fig. 1
EGFP	nkuasgfp1aTg; p188Tg; smb1226Tg	standard conditions	20-25 somites to Prim-15	skeletal muscle cell	IFL	Fig. 1
GFP	ml2Tg	standard conditions	26+ somites to Prim-25	CaP motoneuron axon	IFL	Fig. 1
GFP	ml2Tg	standard conditions	26+ somites to Prim-25	CaP motoneuron	IFL	Fig. 1
GFP	ml2Tg	standard conditions	26+ somites to Prim-25	motor neuron	IFL	Fig. 1
GFP	p187Tg	standard conditions	Prim-5	motor neuron membrane	IHC	Fig. 4
GFP	ppp1r12a^{p82emcf/p82emcf}; p187Tg	standard conditions	20-25 somites to Prim-15	CaP motoneuron axon	IFL	Fig. 5
GFP	ppp1r12a^{p82emcf/p82emcf}; p187Tg	standard conditions	20-25 somites to Prim-15	CaP motoneuron cell body	IFL	Fig. 5
GFP	ppp1r12a^{p82emcf/p82emcf}; p187Tg	standard conditions	20-25 somites to Prim-15	CaP motoneuron	IFL	Fig. 5

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
ppp1r12a^{p82emcf/p82emcf}	standard conditions	Prim-5	CaP motoneuron axon increased branchiness, abnormal	Fig. 3
ppp1r12a^{p82emcf/p82emcf}	standard conditions	Prim-5	CaP motoneuron axon morphology, abnormal	Fig. 3
ppp1r12a^{p82emcf/p82emcf}	standard conditions	Prim-5	CaP motoneuron axonogenesis process quality, abnormal	Fig. 3
ppp1r12a^{p82emcf/p82emcf}	standard conditions	Prim-5	motor neuron axon guidance process quality, abnormal	Fig. 2
ppp1r12a^{p82emcf/p82emcf}; p187Tg	standard conditions	Prim-5	motor neuron membrane ab3-cdh2 labeling decreased amount, abnormal	Fig. 4
ppp1r12a^{p82emcf/p82emcf}; p187Tg; p188Tg	standard conditions	Prim-5	CaP motoneuron cell body mislocalised anteriorly, abnormal	Fig. 5
ppp1r12a^{p82emcf/p82emcf}; p187Tg; p188Tg	standard conditions	Prim-15	CaP motoneuron cell body mislocalised anteriorly, abnormal	Fig. 5
ppp1r12a^{p82emcf/p82emcf}; p188Tg	control	14-19 somites	motor neuron bleb assembly increased occurrence, abnormal	Fig. 4
ppp1r12a^{p82emcf/p82emcf}; p188Tg	control	Prim-5	CaP motoneuron cell body mislocalised anteriorly, abnormal	Fig. 4
ppp1r12a^{p82emcf/p82emcf}; p188Tg	standard conditions	14-19 somites	motor neuron bleb assembly increased occurrence, abnormal	Fig. 4

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
ml2Tg	Tg(mnx1:GFP)	Transgenic Insertion
nkuasgfp1aTg	Tg(5xUAS:EGFP)	Transgenic Insertion
p82emcf		Point Mutation	ppp1r12a
p187Tg	Tg(mnx1:Mmu.Cd8-GFP,mnx1:Mmu.Cd8-GFP)	Transgenic Insertion
p188Tg	Tg(mnx1:mKate2,mnx1:Mmu.Cd8-mKate2)	Transgenic Insertion
smb1226Tg	TgBAC(Tru.Evx1:Tru.Evx1-GAL4FF)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Fish

Fish
ml2Tg
nkuasgfp1aTg; p188Tg; smb1226Tg
p187Tg
p187Tg; p188Tg
p188Tg
ppp1r12a^{p82emcf/p82emcf}
ppp1r12a^{p82emcf/p82emcf}; p187Tg
ppp1r12a^{p82emcf/p82emcf}; p187Tg; p188Tg
ppp1r12a^{p82emcf/p82emcf}; p188Tg
WT

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GAL4FF	EFG	GAL4FF
GFP	EFG	GFP
mKate2	EFG	mKate2

Mapping

Bremer et al., 2016 ZDB-PUB-161118-2 PMID:27855159

Myosin phosphatase Fine-tunes Zebrafish Motoneuron Position during Axonogenesis

Bremer et al., 2016

ZDB-PUB-161118-2
PMID:27855159