PUBLICATION

Live fate-mapping of joint-associated fibroblasts visualizes expansion of cell contributions during zebrafish fin regeneration

Authors

Tornini, V.A., Thompson, J.D., Allen, R.L., Poss, K.D.

ID

ZDB-PUB-170817-6

Date

2017

Source

Development (Cambridge, England) 144: 2889-2895 (Journal)

Registered Authors

Poss, Kenneth D., Thompson, John D., Tornini, Valerie A.

Keywords

Clonal analysis, Fin regeneration, Regeneration, Zebrafish

MeSH Terms

Zebrafish
Animals
Fibroblasts/cytology*
Fibroblasts/metabolism
Joints/metabolism
Joints/physiology
Regeneration/genetics
Regeneration/physiology*
Animal Fins/cytology
Animal Fins/metabolism

(all 10)

PubMed

28811310 Full text @ Development

Abstract

The blastema is a mass of progenitor cells responsible for regeneration of amputated salamander limbs and fish fins. Previous studies have indicated that resident cell sources producing the blastema contribute lineage-restricted progeny to regenerating tissue. However, these studies have labeled general cell types rather than granular cell subpopulations, and they do not explain the developmental transitions that must occur for distal structures to arise from cells with proximal identities in the appendage stump. Here, we find that regulatory sequences of tph1b, which encodes an enzyme that synthesizes serotonin, mark a subpopulation of fibroblast-like cells restricted to the joints of uninjured adult zebrafish fins. Amputation stimulates serotonin production in regenerating fin fibroblasts, yet targeted tph1b mutations abrogating this response do not disrupt fin regeneration. In uninjured animals, tph1b-expressing cells contribute fibroblast progeny that remain restricted to joints throughout life. By contrast, upon amputation, tph1b⁺ joint cells give rise to fibroblasts that distribute across the entire lengths of regenerating fin rays. Our experiments visualize and quantify how incorporation into an appendage blastema broadens the progeny contributions of a cellular subpopulation that normally has proximodistal restrictions.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
EGFP	cz1701Tg; pd250Tg	amputation: caudal fin	Adult	caudal fin blastema	IFL	Fig. 3
EGFP	cz1701Tg; pd250Tg	amputation: caudal fin	Adult	caudal fin fibroblast	IFL	Fig. 3
EGFP	cz1701Tg; pd250Tg	chemical treatment by environment: tamoxifen	Adult	caudal fin lepidotrichium lepidotrichium joint	IFL	Fig. 3
EGFP	ens700Tg; pd51Tg	standard conditions	Adult	caudal fin lepidotrichium osteoblast	IHC	Fig. 1
EGFP	ens700Tg; y1Tg	standard conditions	Adult	caudal fin blood vessel	IFL	Fig. 1
EGFP	evx1^i232/i232; cz1701Tg; pd250Tg	amputation: caudal fin	Adult	caudal fin regenerating fin	IFL	Fig. S1
EGFP	evx1^i232/i232; cz1701Tg; pd250Tg	chemical treatment by environment: tamoxifen	Adult	caudal fin regenerating tissue	IFL	Fig. S1
mCherry	ens700Tg	standard conditions	Adult	anal fin lepidotrichium lepidotrichium joint	IFL	Fig. 1
mCherry	ens700Tg	chemical treatment by environment: tamoxifen, amputation: caudal fin	Adult	caudal fin fibroblast	IFL	Fig. 1
mCherry	ens700Tg	standard conditions	Adult	caudal fin fibroblast	IHC	Fig. 2

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
evx1^i232/i232; ens700Tg/ens700Tg	standard conditions	Days 30-44	caudal fin lepidotrichium broken, abnormal	Fig. 1
evx1^i232/i232; ens700Tg/ens700Tg	standard conditions	Days 30-44	caudal fin lepidotrichium lacks parts or has fewer parts of type lepidotrichium joint, abnormal	Fig. 1
evx1^i232/i232; ens700Tg/ens700Tg	standard conditions	Days 30-44	caudal fin lepidotrichium mCherry expression spatial pattern, abnormal	Fig. 1
evx1^i232/i232; ens700Tg/ens700Tg	standard conditions	Adult	caudal fin deformed, abnormal	Fig. 1
evx1^i232/i232; ens700Tg/ens700Tg	standard conditions	Adult	caudal fin lepidotrichium broken, abnormal	Fig. 1
evx1^i232/i232; ens700Tg/ens700Tg	standard conditions	Adult	caudal fin lepidotrichium lacks parts or has fewer parts of type lepidotrichium joint, abnormal	Fig. 1
evx1^i232/i232; ens700Tg/ens700Tg	standard conditions	Adult	caudal fin lepidotrichium mCherry expression spatial pattern, abnormal	Fig. 1
tph1b^pd249/pd249	standard conditions	Days 45-89	growth decreased efficacy, abnormal	Fig. 2
tph1b^pd249/pd249	standard conditions	Days 45-89	whole organism decreased length, abnormal	Fig. 2
tph1b^pd249/pd249	standard conditions	Days 45-89	whole organism decreased weight, abnormal	Fig. 2

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
cz1701Tg	Tg(-3.5ubb:LOXP-EGFP-LOXP-mCherry)	Transgenic Insertion
dt2		Unknown	lof
ens700Tg	Tg(tph1b:mCherry,myl7:EGFP)	Transgenic Insertion
i232		Point Mutation	evx1
pd51Tg	Tg(Ola.Sp7:EGFP-CAAX)	Transgenic Insertion
pd249		Small Deletion	tph1b
pd250Tg	Tg(tph1b:Cre-ERT2,cryaa:EGFP)	Transgenic Insertion
pd275Tg	Tg(tph1b:mCherry-NTR)	Transgenic Insertion
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
tph1b	CRISPR1-tph1b	CRISPR
tph1b	CRISPR2-tph1b	CRISPR

Fish

Fish
cz1701Tg; pd250Tg
ens700Tg
ens700Tg; pd51Tg
ens700Tg (TL)
ens700Tg; y1Tg
evx1^i232/i232; cz1701Tg; pd250Tg
evx1^i232/i232; ens700Tg/ens700Tg
lof^dt2/dt2; ens700Tg/ens700Tg
pd275Tg
tph1b^pd249/pd249

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab2-serotonin	polyclonal		IgG	Rabbit
zns-5	monoclonal		IgG1	Mouse

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Cre	EFG	Cre
EGFP	EFG	EGFP
mCherry	EFG	mCherry
NTR	EFG	NTR

Mapping

Tornini et al., 2017 ZDB-PUB-170817-6 PMID:28811310

Live fate-mapping of joint-associated fibroblasts visualizes expansion of cell contributions during zebrafish fin regeneration

Tornini et al., 2017

ZDB-PUB-170817-6
PMID:28811310