PUBLICATION

Ablation of EYS in zebrafish causes mislocalisation of outer segment proteins, F-actin disruption and cone-rod dystrophy

Authors

Lu, Z., Hu, X., Liu, F., Soares, D.C., Liu, X., Yu, S., Gao, M., Han, S., Qin, Y., Li, C., Jiang, T., Luo, D., Guo, A.Y., Tang, Z., Liu, M.

ID

ZDB-PUB-170406-5

Date

2017

Source

Scientific Reports 7: 46098 (Journal)

Registered Authors

Gao, Meng, Han, Shanshan, Hu, Xuebin, Li, Chang, Liu, Fei, Liu, Mugen, Liu, Xiliang, Lu, Zhaojing, Qin, Yayun, Yu, Shanshan

Keywords

Development, Retina

MeSH Terms

Rod Cell Outer Segment/metabolism*
Rod Cell Outer Segment/pathology
Rod Cell Outer Segment/ultrastructure
Gene Deletion*
Gene Knockout Techniques
Zebrafish Proteins/metabolism*
Apoptosis
Animals
Light Signal Transduction
Electroretinography
Vision, Ocular
Eye Proteins/metabolism*
Zebrafish/metabolism*
Actins/metabolism*
Base Sequence
Cone-Rod Dystrophies/metabolism*
Cone-Rod Dystrophies/pathology
Cone-Rod Dystrophies/physiopathology
Aging/pathology
Transcription Activator-Like Effector Nucleases/metabolism

(all 20)

PubMed

28378834 Full text @ Sci. Rep.

Abstract

Mutations in EYS are associated with autosomal recessive retinitis pigmentosa (arRP) and autosomal recessive cone-rod dystrophy (arCRD) however, the function of EYS and the molecular mechanisms of how these mutations cause retinal degeneration are still unclear. Because EYS is absent in mouse and rat, and the structure of the retina differs substantially between humans and Drosophila, we utilised zebrafish as a model organism to study the function of EYS in the retina. We constructed an EYS-knockout zebrafish-line by TALEN technology which showed visual impairment at an early age, while the histological and immunofluorescence assays indicated the presence of progressive retinal degeneration with a cone predominately affected pattern. These phenotypes recapitulate the clinical manifestations of arCRD patients. Furthermore, the EYS^-/- zebrafish also showed mislocalisation of certain outer segment proteins (rhodopsin, opn1lw, opn1sw1, GNB3 and PRPH2), and disruption of actin filaments in photoreceptors. Protein mislocalisation may, therefore, disrupt the function of cones and rods in these zebrafish and cause photoreceptor death. Collectively, these results point to a novel role for EYS in maintaining the morphological structure of F-actin and in protein transport, loss of this function might be the trigger for the resultant cellular events that ultimately lead to photoreceptor death.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
gnb1a	eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
gnb1a	WT	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
gnb1b	eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
gnb1b	WT	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
gnb3a	eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
gnb3a	WT	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
gnb3b	eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
gnb3b	WT	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
grk1a	eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2
grk1a	WT	standard conditions	Days 7-13	whole organism	RTPCR	Fig. 2

1 - 10 of 26

Show

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
eys^hzu4/hzu4	standard conditions	Days 7-13	retina photoreceptor inner segment Ab2-opn1lw labeling mislocalised, abnormal	Fig. 7
eys^hzu4/hzu4	standard conditions	Days 7-13	retina photoreceptor inner segment Ab2-opn1sw2 labeling mislocalised, abnormal	Fig. 7
eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism gnb1b expression decreased amount, abnormal	Fig. 2
eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism grk7a expression decreased amount, abnormal	Fig. 2
eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism guca1aa expression decreased amount, abnormal	Fig. 2
eys^hzu4/hzu4	standard conditions	Days 7-13	whole organism pde6c expression decreased amount, abnormal	Fig. 2
eys^hzu4/hzu4	standard conditions	Days 30-44	retinal outer nuclear layer thickness, normal	Fig. 3
eys^hzu4/hzu4	standard conditions	Days 45-89	retina apoptotic process increased occurrence, abnormal	Fig. 3
eys^hzu4/hzu4	standard conditions	Days 45-89	retina apoptotic process occurrence, normal	Fig. S4
eys^hzu4/hzu4	standard conditions	Days 45-89	retinal outer nuclear layer decreased thickness, abnormal	Fig. 3

1 - 10 of 31

Show

Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
hzu4		Small Deletion	eys

Human Disease / Model

Human Disease	Fish	Conditions	Evidence
cone-rod dystrophy	eys^hzu4/hzu4	standard conditions	TAS
retinitis pigmentosa			TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
eys	TALEN1-eys	TALEN

Fish

Fish
eys^hzu4/hzu4
WT

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-eys	polyclonal	eys	IgG	Rabbit
Ab1-gnb3	polyclonal		IgG	Rabbit
Ab1-prph2	polyclonal		IgG	Rabbit
Ab2-opn1lw
Ab2-opn1sw2	polyclonal		IgG	Rabbit

Orthology

Engineered Foreign Genes

Mapping

Lu et al., 2017 ZDB-PUB-170406-5 PMID:28378834

Ablation of EYS in zebrafish causes mislocalisation of outer segment proteins, F-actin disruption and cone-rod dystrophy

Lu et al., 2017

ZDB-PUB-170406-5
PMID:28378834