PUBLICATION

Orphan G-Protein Coupled Receptor 22 (Gpr22) Regulates Cilia Length and Structure in the Zebrafish Kupffer's Vesicle

Authors

Verleyen, D., Luyten, F.P., Tylzanowski, P.

ID

ZDB-PUB-141022-1

Date

2014

Source

PLoS One 9: e110484 (Journal)

Registered Authors

Keywords

none

MeSH Terms

Zebrafish Proteins/antagonists & inhibitors
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
RNA Interference
Phenotype
Microscopy, Electron, Transmission
Transcription Factors/metabolism
Forkhead Transcription Factors/metabolism
Receptors, G-Protein-Coupled/antagonists & inhibitors
Receptors, G-Protein-Coupled/genetics
Receptors, G-Protein-Coupled/metabolism*
Body Patterning
Oligonucleotides, Antisense/metabolism
Cilia/chemistry
Cilia/physiology*
Embryo, Nonmammalian/metabolism
Zebrafish/growth & development
Animals
Embryonic Development/physiology
RNA, Messenger/metabolism

(all 20)

PubMed

25335082 Full text @ PLoS One

Abstract

GPR22 is an orphan G protein-coupled receptor (GPCR). Since the ligand of the receptor is currently unknown, its biological function has not been investigated in depth. Many GPCRs and their intracellular effectors are targeted to cilia. Cilia are highly conserved eukaryotic microtubule-based organelles that protrude from the membrane of most mammalian cells. They are involved in a large variety of physiological processes and diseases. However, the details of the downstream pathways and mechanisms that maintain cilia length and structure are poorly understood. We show that morpholino knock down or overexpression of gpr22 led to defective left-right (LR) axis formation in the zebrafish embryo. Specifically, defective LR patterning included randomization of the left-specific lateral plate mesodermal genes (LPM) (lefty1, lefty2, southpaw and pitx2a), resulting in randomized cardiac looping. Furthermore, gpr22 inactivation in the Kupffer's vesicle (KV) alone was still able to generate the phenotype, indicating that Gpr22 mainly regulates LR asymmetry through the KV. Analysis of the KV cilia by immunofluorescence and transmission electron microscopy (TEM), revealed that gpr22 knock down or overexpression resulted in changes of cilia length and structure. Further, we found that Gpr22 does not act upstream of the two cilia master regulators, Foxj1a and Rfx2. To conclude, our study characterized a novel player in the field of ciliogenesis.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
dand5	WT	control	10-13 somites	Kupffer's vesicle	ISH	Fig. 4
dand5	WT + MO1-gpr22a + MO4-tp53	standard conditions	10-13 somites	Kupffer's vesicle	ISH	Fig. 4
EGFP	twu34Tg	control	Day 4	heart	IFL	Fig. 2
EGFP	twu34Tg + MO1-gpr22a + MO4-tp53	standard conditions	Day 4	heart	IFL	Fig. 2
foxa3	WT	control	Day 4	gut	ISH	Fig. 2
foxa3	WT	control	Day 4	liver	ISH	Fig. 2
foxa3	WT + MO1-gpr22a + MO4-tp53	standard conditions	Day 4	gut	ISH	Fig. 2
foxa3	WT + MO1-gpr22a + MO4-tp53	standard conditions	Day 4	liver	ISH	Fig. 2
foxj1a	WT	control	Bud	forerunner cell group	ISH	Fig. 6
foxj1a	WT + MO1-gpr22a + MO4-tp53	standard conditions	Bud	forerunner cell group	ISH	Fig. 6

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + MO1-gpr22a + MO4-tp53	standard conditions	10-13 somites	Kupffer's vesicle cilium decreased length, abnormal	Fig. 5
WT + MO1-gpr22a + MO4-tp53	standard conditions	10-13 somites	Kupffer's vesicle cilium disorganized, abnormal	Fig. 5
WT + MO1-gpr22a + MO4-tp53	standard conditions	10-13 somites	Kupffer's vesicle cilium has fewer parts of type Kupffer's vesicle axonemal microtubule, abnormal	Fig. 5
WT + MO1-gpr22a + MO4-tp53	standard conditions	20-25 somites	determination of left/right asymmetry in lateral mesoderm process quality, abnormal	Fig. 3
WT + MO1-gpr22a + MO4-tp53	standard conditions	20-25 somites	determination of left/right asymmetry in lateral mesoderm process quality, abnormal	Fig. 2
WT + MO1-gpr22a + MO4-tp53	standard conditions	20-25 somites	left/right axis specification process quality, abnormal	Fig. 2
WT + MO1-gpr22a + MO4-tp53	standard conditions	Prim-5	caudal fin curved ventral, abnormal	Fig. 1
WT + MO1-gpr22a + MO4-tp53	standard conditions	Prim-5	heart edematous, abnormal	Fig. 1
WT + MO1-gpr22a + MO4-tp53	standard conditions	Prim-5	whole organism decreased length, abnormal	Fig. 1
WT + MO1-gpr22a + MO4-tp53	standard conditions	Long-pec	inner ear has extra parts of type otolith, abnormal	Fig. S6

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
twu34Tg	Tg(myl7:EGFP)	Transgenic Insertion

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
gpr22a	MO1-gpr22a	MRPHLNO
gpr22a	MO2-gpr22a	MRPHLNO
tp53	MO4-tp53	MRPHLNO

Fish

Fish
twu34Tg
twu34Tg + MO1-gpr22a + MO4-tp53
WT
WT + MO1-gpr22a + MO4-tp53

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP

Mapping

Verleyen et al., 2014 ZDB-PUB-141022-1 PMID:25335082

Orphan G-Protein Coupled Receptor 22 (Gpr22) Regulates Cilia Length and Structure in the Zebrafish Kupffer's Vesicle

Verleyen et al., 2014

ZDB-PUB-141022-1
PMID:25335082