PUBLICATION

Distinct requirements of wls, wnt9a, wnt5b and gpc4 in regulating chondrocyte maturation and timing of endochondral ossification

Authors

Ling, I.T., Rochard, L., Liao, E.C.

ID

ZDB-PUB-161203-3

Date

2017

Source

Developmental Biology 421(2): 219-232 (Journal)

Registered Authors

Liao, Eric

Keywords

Meckel's cartilage, Wnt, chondrocyte, endochondral ossification, osteoblast, zebrafish

MeSH Terms

Face
Wnt Proteins/metabolism*
Gene Expression Regulation, Developmental
Chondrogenesis*
Cell Polarity
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Osteogenesis*
Zebrafish
Cartilage/metabolism
Cartilage/pathology
Models, Biological
Animals
Cell Differentiation*
Muscles/metabolism
Muscles/pathology
Joints/metabolism
Joints/pathology
Mutation/genetics
Chondrocytes/cytology*
Chondrocytes/metabolism
Skull/metabolism
Time Factors
Cell Proliferation
Wnt Signaling Pathway

(all 25)

PubMed

27908786 Full text @ Dev. Biol.

Abstract

Formation of the mandible requires progressive morphologic change, proliferation, differentiation and organization of chondrocytes preceding osteogenesis. The Wnt signaling pathway is involved in regulating bone development and maintenance. Chondrocytes that are fated to become bone require Wnt to polarize and orientate appropriately to initiate the endochondral ossification program. Although the canonical Wnt signaling has been well studied in the context of bone development, the effects of non-canonical Wnt signaling in regulating the timing of cartilage maturation and subsequent bone formation in shaping ventral craniofacial structure is not fully understood.. Here we examined the role of the non-canonical Wnt signaling pathway (wls, gpc4, wnt5b and wnt9a) in regulating zebrafish Meckel's cartilage maturation to the onset of osteogenic differentiation. We found that disruption of wls resulted in a significant loss of craniofacial bone, whereas lack of gpc4, wnt5b and wnt9a resulted in severely delayed endochondral ossification. This study demonstrates the importance of the non-canonical Wnt pathway in regulating coordinated ventral cartilage morphogenesis and ossification.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
col10a1a	WT	standard conditions	Day 4	ceratobranchial 5 cartilage	ISH	Fig. 7
col10a1a	WT	standard conditions	Day 4	ceratohyal cartilage	ISH	Fig. 7
col10a1a	WT	standard conditions	Day 4	cleithrum	ISH	Fig. 7
col10a1a	WT	standard conditions	Day 4	Meckel's cartilage	ISH	Fig. 7
col1a2	WT	standard conditions	Day 4	Meckel's cartilage	ISH	Fig. 7
col2a1a	gpc4^{hi1688Tg/hi1688Tg}	standard conditions	Day 4	Meckel's cartilage	ISH	Fig. 7
col2a1a	wls^c186/c186	standard conditions	Day 4	Meckel's cartilage	ISH	Fig. 7
col2a1a	wnt5b^{hi2735bTg/hi2735bTg}	standard conditions	Day 4	Meckel's cartilage	ISH	Fig. 7
col2a1a	wnt9a^zf754/zf754	standard conditions	Day 4	Meckel's cartilage	ISH	Fig. 7
col2a1a	WT	standard conditions	Day 4	Meckel's cartilage	ISH	Fig. 7

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
gpc4^{hi1688Tg/hi1688Tg}	standard conditions	Day 4	chondrocyte amount, normal	Fig. 2
gpc4^{hi1688Tg/hi1688Tg}	standard conditions	Day 4	Meckel's cartilage shortened, abnormal	Fig. 2
gpc4^{hi1688Tg/hi1688Tg}	standard conditions	Days 7-13	interhyal cartilage chondrocyte decreased amount, abnormal	Fig. 5
gpc4^{hi1688Tg/hi1688Tg}	standard conditions	Days 7-13	quadrate-anguloarticular joint morphology, abnormal	Fig. 5
gpc4^{hi1688Tg/hi1688Tg}	standard conditions	Days 7-13	retroarticular malformed, abnormal	Fig. 5
gpc4^{hi1688Tg/hi1688Tg}	standard conditions	Days 7-13	retroarticular shortened, abnormal	Fig. 5
gpc4^{hi1688Tg/hi1688Tg}; ir937Tg	standard conditions	Protruding-mouth	chondrocyte microtubule organizing center disorganized, abnormal	Fig. 3
gpc4^{hi1688Tg/hi1688Tg}; ir937Tg	standard conditions	Protruding-mouth	Meckel's cartilage cartilage morphogenesis process quality, abnormal	Fig. 3
gpc4^{hi1688Tg/hi1688Tg}; ir937Tg	standard conditions	Protruding-mouth	Meckel's cartilage chondrocyte morphology, abnormal	Fig. 3
gpc4^{hi1688Tg/hi1688Tg}; ir937Tg	standard conditions	Protruding-mouth	Meckel's cartilage chondrocyte positional polarity, abnormal	Fig. 3

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
c186		Point Mutation	wls
hi1688Tg	Tg(nLacz-GTvirus)	Transgenic Insertion	gpc4
hi2735bTg	Tg(nLacz-GTvirus)	Transgenic Insertion	wnt5b
ir937Tg	Tg(-7.2sox10:EGFP)	Transgenic Insertion
pd44Tg	Tg2(Ola.Bglap:EGFP)	Transgenic Insertion
pd46Tg	Tg(Ola.Sp7:mCherry-Eco.NfsB)	Transgenic Insertion
zf595Tg	Tg(sox10:mCherry)	Transgenic Insertion
zf754		Small Deletion	wnt9a

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
wnt9a	CRISPR2-wnt9a	CRISPR

Fish

Fish
gpc4^{hi1688Tg/hi1688Tg}
gpc4^{hi1688Tg/hi1688Tg}; ir937Tg
ir937Tg; zf595Tg
wls^c186/c186
wls^c186/c186; ir937Tg
wnt5b^{hi2735bTg/hi2735bTg}
wnt5b^{hi2735bTg/hi2735bTg}; ir937Tg
wnt9a^zf754/zf754
wnt9a^zf754/zf754; ir937Tg
WT

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
mCherry	EFG	mCherry

Mapping

Ling et al., 2017 ZDB-PUB-161203-3 PMID:27908786

Distinct requirements of wls, wnt9a, wnt5b and gpc4 in regulating chondrocyte maturation and timing of endochondral ossification

Ling et al., 2017

ZDB-PUB-161203-3
PMID:27908786