PUBLICATION

Hsp47 mediates Cx43-dependent skeletal growth and patterning in the regenerating fin

Authors

Bhadra, J., Iovine, M.K.

ID

ZDB-PUB-150624-1

Date

2015

Source

Mechanisms of Development 138 Pt 3: 364-74 (Journal)

Registered Authors

Iovine, M. Kathryn

Keywords

cell proliferation, gap junction, joint formation, short fin, zebrafish

MeSH Terms

Models, Biological
Body Patterning/genetics
Body Patterning/physiology
Zebrafish Proteins/antagonists & inhibitors
Zebrafish Proteins/genetics
Zebrafish Proteins/physiology*
Regeneration/genetics
Regeneration/physiology
RNA, Messenger/genetics
RNA, Messenger/metabolism
Zebrafish/genetics
Zebrafish/growth & development*
Zebrafish/physiology*
Gene Knockdown Techniques
Mutation
Animals, Genetically Modified
Animal Fins/growth & development*
Animal Fins/physiology*
HSP47 Heat-Shock Proteins/antagonists & inhibitors
HSP47 Heat-Shock Proteins/genetics
HSP47 Heat-Shock Proteins/physiology*
Signal Transduction
Bone Development/genetics
Bone Development/physiology
Cell Proliferation
Animals
In Situ Hybridization
Connexin 43/genetics
Connexin 43/physiology*

(all 29)

PubMed

26103547 Full text @ Mech. Dev.

Abstract

Skeletal morphogenesis describes how bones achieve their correct shape and size and appropriately position joints. We use the regenerating caudal fin of zebrafish to study this process. Our examination of the fin length mutant short fin (sof (b123)) has revealed that the gap junction protein Cx43 is involved in skeletal morphogenesis by promoting cell proliferation and inhibiting joint formation, thereby coordinating skeletal growth and patterning. Here we demonstrate that serpinh1b is molecularly and functionally downstream of cx43. The gene serpinh1b codes for a protein called Hsp47, a molecular chaperone responsible for proper folding of procollagen molecules. Knockdown of Hsp47 in regenerating fins recapitulates the sof (b123) phenotypes of reduced fin length, reduced segment length and reduced level of cell proliferation. Furthermore, Hsp47 knockdown affects the organization and localization of the collagen-based actinotrichia. Together, our findings reveal that serpinh1b acts in a cx43 dependent manner to regulate cell proliferation and joint formation. We conclude that disruption of the collagen-based extracellular matrix influences signaling events required for cell proliferation as well as the patterning of skeletal precursor cells that influences segment length. Therefore, we suggest that Hsp47 function is necessary for skeletal growth and patterning during fin regeneration.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
msx1b	WT + MO1-serpinh1b	amputation: fin	Adult	regenerating fin	RTPCR	Fig. 8
msx3	WT + MO1-serpinh1b	amputation: fin	Adult	regenerating fin	RTPCR	Fig. 8
serpinh1b	gja1b^b123/b123	amputation: fin	Adult	regenerating fin	RTPCR	Fig. 2
serpinh1b	gja1b^b123/b123	amputation: fin	Adult	regenerating fin	ISH	Fig. 1
serpinh1b	kcnk5b^ty86d/ty86d	amputation: fin	Adult	regenerating fin	RTPCR	Fig. 2
serpinh1b	kcnk5b^ty86d/ty86d	amputation: fin	Adult	regenerating fin	ISH	Fig. 1
serpinh1b	WT	amputation: fin	Adult	basal regeneration epithelium	ISH	Fig. 1
serpinh1b	WT	amputation: fin	Adult	basal regeneration epithelium	ISH	Fig. 3
serpinh1b	WT	amputation: fin	Adult	blastema	ISH	Fig. 1
serpinh1b	WT	amputation: fin	Adult	blastema	ISH	Fig. 3

1 - 10 of 16

Show

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + MO1-hapln1a	amputation: fin	Adult	regenerating fin serpinh1b expression decreased amount, abnormal	Fig. 2
WT + MO1-serpinh1b	standard conditions	Adult	fin regeneration decreased efficacy, abnormal	Fig. 5
WT + MO1-serpinh1b	standard conditions	Adult	fin regeneration decreased process quality, abnormal	Fig. 5
WT + MO1-serpinh1b	standard conditions	Adult	regenerating fin cell population proliferation decreased occurrence, abnormal	Fig. 5
WT + MO1-serpinh1b	standard conditions	Adult	regenerating fin decreased length, abnormal	Fig. 5
WT + MO1-serpinh1b	amputation: fin	Adult	lepidotrichium joint fin morphogenesis premature, abnormal	Fig. 7
WT + MO1-serpinh1b	amputation: fin	Adult	lepidotrichium joint fin regeneration premature, abnormal	Fig. 7
WT + MO1-serpinh1b	amputation: fin	Adult	regenerating fin msx1b expression decreased amount, abnormal	Fig. 8
WT + MO1-serpinh1b	amputation: fin	Adult	regenerating fin msx3 expression decreased amount, abnormal	Fig. 8
WT + MO1-serpinh1b	amputation: fin	Adult	regenerating fin actinotrichium decreased size, abnormal	Fig. 11

1 - 10 of 24

Show

Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
b123		Unknown	gja1b
ty86d		Point Mutation	kcnk5b

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
gja1b	MO2-gja1b	MRPHLNO
hapln1a	MO1-hapln1a	MRPHLNO
sema3d	MO5-sema3d	MRPHLNO
serpinh1b	MO1-serpinh1b	MRPHLNO

Fish

Fish
gja1b^b123/b123
kcnk5b^ty86d/ty86d
WT
WT + MO1-hapln1a
WT + MO1-serpinh1b
WT + MO2-gja1b

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Bhadra et al., 2015 ZDB-PUB-150624-1 PMID:26103547

Hsp47 mediates Cx43-dependent skeletal growth and patterning in the regenerating fin

Bhadra et al., 2015

ZDB-PUB-150624-1
PMID:26103547