PUBLICATION

The ubiquitin ligase PHR promotes directional regrowth of spinal zebrafish axons

Authors

Bremer, J., Marsden, K.C., Miller, A., Granato, M.

ID

ZDB-PUB-190604-1

Date

2019

Source

Communications biology 2: 195 (Journal)

Registered Authors

Granato, Michael, Marsden, Kurt, Miller, Adam

Keywords

Spinal cord diseases, Spinal cord injury

MeSH Terms

Actins/metabolism
Cytoplasmic Dyneins/metabolism
Alleles
Ubiquitin/metabolism
Image Processing, Computer-Assisted
Cadherins/metabolism
Adaptor Proteins, Signal Transducing/metabolism*
MAP Kinase Kinase 4/metabolism
Mixed Function Oxygenases/metabolism*
Transgenes
Green Fluorescent Proteins/metabolism
Neuronal Outgrowth*
Animals
Zebrafish Proteins/metabolism*
Axons/metabolism*
Spinal Cord Injuries/metabolism
Mutation
Animals, Genetically Modified
Ligases/metabolism
Spinal Cord/pathology
Disease Models, Animal
Nerve Regeneration*
Zebrafish

(all 23)

PubMed

31149640 Full text @ Commun Biol

Abstract

To reconnect with their synaptic targets, severed axons need to regrow robustly and directionally along the pre-lesional trajectory. While mechanisms directing axonal regrowth are poorly understood, several proteins direct developmental axon outgrowth, including the ubiquitin ligase PHR (Mycbp2). Invertebrate PHR also limits regrowth of injured axons, whereas its role in vertebrate axonal regrowth remains elusive. Here we took advantage of the high regrowth capacity of spinal zebrafish axons and observed robust and directional regrowth following laser transection of spinal Mauthner axons. We found that PHR directs regrowing axons along the pre-lesional trajectory and across the transection site. At the transection site, initial regrowth of wild-type axons was multidirectional. Over time, misdirected sprouts were corrected in a PHR-dependent manner. Ablation of cyfip2, known to promote F-actin-polymerization and pharmacological inhibition of JNK reduced misdirected regrowth of PHR-deficient axons, suggesting that PHR controls directional Mauthner axonal regrowth through cyfip2- and JNK-dependent pathways.

Genes / Markers

Marker	Marker Type	Name
celsr3	GENE	cadherin, EGF LAG seven-pass G-type receptor 3
cfl1l	GENE	cofilin 1 (non-muscle), like
cyfip2	GENE	cytoplasmic FMR1 interacting protein 2
dgcr8	GENE	DGCR8 microprocessor complex subunit
dicer1	GENE	dicer 1, ribonuclease type III
dync1h1	GENE	dynein, cytoplasmic 1, heavy chain 1
gfap	GENE	glial fibrillary acidic protein
lrp4	GENE	low density lipoprotein receptor-related protein 4
mycbp2	GENE	MYC binding protein 2
nf1a	GENE	neurofibromin 1a

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Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
Citrine	nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 6 to Days 7-13	Mauthner neuron axon	IFL	Fig. 1
Citrine	nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 6 to Days 7-13	Mauthner neuron	IFL	Fig. 1
EGFP	mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p411Tg; ue2Tg	transection: Mauthner neuron	Day 5 to Days 7-13	oligodendrocyte	IFL	Fig. 6
EGFP	nkhspGFF62ATg; p411Tg; ue2Tg	transection: Mauthner neuron	Day 5	oligodendrocyte	IFL	Fig. 1
EGFP	nkhspGFF62ATg; p411Tg; ue2Tg	transection: Mauthner neuron	Day 5 to Days 7-13	oligodendrocyte	IFL	Fig. 6
GFP	mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p411Tg; zf167Tg	transection: Mauthner neuron	Day 5 to Days 7-13	glial cell	IFL	Fig. 6
GFP	nkhspGFF62ATg; p411Tg; zf167Tg	transection: Mauthner neuron	Day 5	glial cell	IFL	Fig. 1
GFP	nkhspGFF62ATg; p411Tg; zf167Tg	transection: Mauthner neuron	Day 5 to Days 7-13	glial cell	IFL	Fig. 6
RFP	mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p411Tg; ue2Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon	IFL	Fig. 6
RFP	mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p411Tg; ue2Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron	IFL	Fig. 6

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
celsr3^fh339/fh339; nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon extension involved in regeneration decreased efficacy, abnormal	Fig. 2
celsr3^fh339/fh339; nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon extension involved in regeneration process quality, abnormal	Fig. 2
celsr3^fh339/fh339; nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon regeneration decreased efficacy, abnormal	Fig. 2
celsr3^fh339/fh339; nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon regeneration process quality, abnormal	Fig. 2
cyfip2^p400/p400; mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p201Tg	standard conditions	Days 7-13	Mauthner neuron axon extension involved in regeneration process quality, ameliorated	Fig. 8
cyfip2^p400/p400; mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p201Tg	standard conditions	Days 7-13	Mauthner neuron axon regeneration process quality, abnormal	Fig. 8
cyfip2^p400/p400; nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon extension involved in regeneration decreased efficacy, abnormal	Fig. 2
cyfip2^p400/p400; nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon extension involved in regeneration process quality, abnormal	Fig. 2
cyfip2^p400/p400; nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon regeneration decreased efficacy, abnormal	Fig. 2
cyfip2^p400/p400; nkhspGFF62ATg; p201Tg	transection: Mauthner neuron	Day 5 to Days 7-13	Mauthner neuron axon regeneration process quality, abnormal	Fig. 2

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
fh339		Point Mutation	celsr3
fh344		Point Mutation	dgcr8
hi3684Tg	Tg(nLacz-GTvirus)	Transgenic Insertion	dync1h1
hu896		Point Mutation	dicer1
m241		Unknown	sox10
ml2Tg	Tg(mnx1:GFP)	Transgenic Insertion
nkhspGFF62ATg	Et(hsp70l:GAL4FF)	Transgenic Insertion
p170		Point Mutation	pappaa
p174		Point Mutation	zdhhc17
p184		Small Deletion	lrp4

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Human Disease / Model

Sequence Targeting Reagents

Fish

Fish
celsr3^fh339/fh339; nkhspGFF62ATg; p201Tg
cyfip2^p400/p400; mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p201Tg
cyfip2^p400/p400; nkhspGFF62ATg; p201Tg
dync1h1^{hi3684Tg/hi3684Tg}; nkhspGFF62ATg; p201Tg
mycbp2^tn207b/+; mycbp2^tp203/+; nkhspGFF62ATg; p201Tg
mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p201Tg
mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p411Tg; ue2Tg
mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p411Tg; zf167Tg
mycbp2^{tn207b/tn207b}; nkhspGFF62ATg; p412Tg
nkhspGFF62ATg; p201Tg

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Citrine	EFG	Citrine
EGFP	EFG	EGFP
GAL4FF	EFG	GAL4FF
GFP	EFG	GFP
RFP	EFG	RFP

Mapping

Bremer et al., 2019 ZDB-PUB-190604-1 PMID:31149640

The ubiquitin ligase PHR promotes directional regrowth of spinal zebrafish axons

Bremer et al., 2019

ZDB-PUB-190604-1
PMID:31149640