PUBLICATION

miR-196 regulates axial patterning and pectoral appendage initiation

Authors

He, X., Yan, Y.L., Eberhart, J.K., Herpin, A., Wagner, T.U., Schartl, M., and Postlethwait, J.H.

ID

ZDB-PUB-110803-10

Date

2011

Source

Developmental Biology 357(2): 463-77 (Journal)

Registered Authors

Eberhart, Johann, He, Xinjun, Postlethwait, John H., Schartl, Manfred, Wagner, Toni, Yan, Yi-Lin

Keywords

microRNA (miRNA), Mir196 (miR-196), axial skeletal patterning, pectoral fin, pharyngeal arch, retinoic acid

MeSH Terms

Animals
Zebrafish/embryology*
Zebrafish/genetics*
Signal Transduction/genetics
Genomics
MicroRNAs/genetics
MicroRNAs/metabolism*
Molecular Sequence Data
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Body Patterning/genetics*
Branchial Region/embryology
Branchial Region/metabolism
Bone and Bones/embryology
Bone and Bones/metabolism
Base Sequence
Animal Fins/embryology*
Animal Fins/metabolism
Gene Expression Regulation, Developmental
Embryo, Nonmammalian/metabolism
Tretinoin/metabolism
Homeodomain Proteins/genetics
Homeodomain Proteins/metabolism

(all 23)

PubMed

21787766 Full text @ Dev. Biol.

Abstract

Vertebrate Hox clusters contain protein-coding genes that regulate body axis development and microRNA (miRNA) genes whose functions are not yet well understood. We overexpressed the Hox cluster microRNA miR-196 in zebrafish embryos and found four specific, viable phenotypes: failure of pectoral fin bud initiation, deletion of the 6th pharyngeal arch, homeotic aberration and loss of rostral vertebrae, and reduced number of ribs and somites. Reciprocally, miR-196 knockdown evoked an extra pharyngeal arch, extra ribs, and extra somites, confirming endogenous roles of miR-196. miR-196 injection altered expression of hox genes and the signaling of retinoic acid through the retinoic acid receptor gene rarab. Knocking down rarab mimicked the pectoral fin phenotype of miR-196 overexpression, and reporter constructs tested in tissue culture and in embryos showed that the rarab 32UTR is a miR-196 target for pectoral fin bud initiation. These results show that a Hox cluster microRNA modulates development of axial patterning similar to nearby protein-coding Hox genes, and acts on appendicular patterning at least in part by modulating retinoic acid signaling.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
EYFP	ld1Tg	standard conditions	High-pec		central nervous system	IFL	Fig. 4
EYFP	ld1Tg + MO1-mir196a-1,mir196a-2 + MO1-mir196b	standard conditions	High-pec		central nervous system	IFL	Fig. 4
EYFP	ld1Tg + MO1-mir196a-1,mir196a-2 + MO1-mir196b	standard conditions	High-pec		whole organism	IFL	Fig. S4
fgf10a	AB/TU	control	20-25 somites		lateral line primordium	ISH	Fig. 4
fgf10a	AB/TU	control	20-25 somites		otic vesicle	ISH	Fig. 4
fgf10a	AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Prim-5		otic vesicle	ISH	Fig. S5
fgf10a	AB/TU	control	Prim-5		otic vesicle	ISH	Fig. S5
fgf10a	AB/TU	control	Prim-5		pectoral fin bud	ISH	Fig. S5
fgf10a	AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Prim-5	Not Detected	pectoral fin bud	ISH	Fig. S5
fgf10a	AB/TU	control	20-25 somites		pectoral fin field	ISH	Fig. 4

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	20-25 somites	pectoral fin field fgf24 expression absent, abnormal	Fig. S5
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Prim-5	pectoral fin bud fgf10a expression absent, abnormal	Fig. S5
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Prim-5	pectoral fin bud fgf24 expression absent, abnormal	Fig. S5
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Prim-5	pectoral fin bud tbx5a expression absent, abnormal	Fig. S5
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	High-pec	pectoral fin absent, abnormal	Fig. 4
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	High-pec	pectoral fin development disrupted, abnormal	Fig. 4
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Long-pec	pectoral fin lbx1a expression absent, abnormal	Fig. S5
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Day 5	pectoral fin absent, abnormal	Fig. S5
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Day 5	pectoral fin development disrupted, abnormal	Fig. S5
AB/TU	chemical treatment by environment: 4-(diethylamino)benzaldehyde	Days 45-89	pectoral fin absent, abnormal	Fig. S5

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
ld1Tg	Tg(RARE-gata2a:NLS-EYFP)	Transgenic Insertion
rw716		Point Mutation	cyp26a1
y1Tg	Tg(fli1:EGFP)	Transgenic Insertion

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
mir196a-1	MO1-mir196a-1,mir196a-2	MRPHLNO
mir196a-2	MO1-mir196a-1,mir196a-2	MRPHLNO
mir196b	MO1-mir196b	MRPHLNO
rarab	MO1-rarab	MRPHLNO

Fish

Fish
AB/TU
AB/TU + MO1-mir196a-1,mir196a-2 + MO1-mir196b
AB/TU + MO1-rarab
cyp26a1^rw716/rw716
ld1Tg
ld1Tg + MO1-mir196a-1,mir196a-2 + MO1-mir196b
y1Tg
y1Tg + MO1-mir196a-1,mir196a-2 + MO1-mir196b

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
EYFP	EFG	EYFP
YFP	EFG	YFP

Mapping

He et al., 2011 ZDB-PUB-110803-10 PMID:21787766

miR-196 regulates axial patterning and pectoral appendage initiation

He et al., 2011

ZDB-PUB-110803-10
PMID:21787766