PUBLICATION

Deletion of Pr130 Interrupts Cardiac Development in Zebrafish

Authors

Yang, J., Li, Z., Gan, X., Zhai, G., Gao, J., Xiong, C., Qiu, X., Wang, X., Yin, Z., Zheng, F.

ID

ZDB-PUB-161116-2

Date

2016

Source

International Journal of Molecular Sciences 17(11): (Journal)

Registered Authors

Yin, Zhan, Zheng, Fang

Keywords

CRISPR-Cas9, cardiac development, pr130, zebrafish

MeSH Terms

Myocardium/enzymology
Myocardium/pathology
Animals
Zebrafish
Apoptosis
Myocytes, Cardiac/enzymology
Heart/embryology*
Protein Phosphatase 2/genetics*
Gene Deletion
Myocardial Contraction

(all 10)

PubMed

27845735 Full text @ Int. J. Mol. Sci.

Abstract

Protein phosphatase 2 regulatory subunit B, alpha (PPP2R3A), a regulatory subunit of protein phosphatase 2A (PP2A), is a major serine/threonine phosphatase that regulates crucial function in development and growth. Previous research has implied that PPP2R3A was involved in heart failure, and PR130, the largest transcription of PPP2R3A, functioning in the calcium release of sarcoplasmic reticulum (SR), plays an important role in the excitation-contraction (EC) coupling. To obtain a better understanding of PR130 functions in myocardium and cardiac development, two pr130-deletion zebrafish lines were generated using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated proteins (Cas) system. Pr130-knockout zebrafish exhibited cardiac looping defects and decreased cardiac function (decreased fractional area and fractional shortening). Hematoxylin and eosin (H&E) staining demonstrated reduced cardiomyocytes. Subsequent transmission electron microscopy revealed that the bright and dark bands were narrowed and blurred, the Z- and M-lines were fogged, and the gaps between longitudinal myocardial fibers were increased. Additionally, increased apoptosis was observed in cardiomyocyte in pr130-knockout zebrafish compared to wild-type (WT). Taken together, our results suggest that pr130 is required for normal myocardium formation and efficient cardiac contractile function.

Genes / Markers

Figures

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Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
ppp2r3a^zf747/zf747	standard conditions	Long-pec	heart decreased functionality, abnormal	Fig. 4
ppp2r3a^zf747/zf747	standard conditions	Long-pec	heart looping disrupted, abnormal	Fig. 3
ppp2r3a^zf747/zf747	standard conditions	Long-pec	heart morphology, abnormal	Fig. 3
ppp2r3a^zf747/zf747	standard conditions	Days 7-13	pericardium edematous, abnormal	Fig. 3
ppp2r3a^zf747/zf747	standard conditions	Days 7-13	whole organism dead, abnormal	Fig. 3
ppp2r3a^zf747/zf747	standard conditions	Days 7-13	whole organism decreased life span, abnormal	Fig. 3
ppp2r3a^zf747/zf747	standard conditions	Adult	cardiac muscle cell decreased amount, abnormal	Fig. 5
ppp2r3a^zf747/zf747	standard conditions	Adult	heart apoptotic process increased occurrence, abnormal	Fig. 7
ppp2r3a^zf747/zf747	standard conditions	Adult	myocardium disorganized, abnormal	Fig. 6
ppp2r3a^zf748/zf748	standard conditions	Long-pec	heart decreased functionality, abnormal	Fig. 4

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
f1Tg	Tg(myl7:GFP)	Transgenic Insertion
zf747		Indel	ppp2r3a
zf748		Small Deletion	ppp2r3a

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
ppp2r3a	CRISPR1-ppp2r3a	CRISPR

Fish

Fish
ppp2r3a^zf747/zf747
ppp2r3a^zf748/zf748
WT

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-ppp2r3a	polyclonal		IgG	Rabbit
Ab13-gapdh	monoclonal		IgG2b	Mouse

Orthology

Gene	Orthology
ppp2r3a

Engineered Foreign Genes

Marker	Marker Type	Name
GFP	EFG	GFP

Mapping

Yang et al., 2016 ZDB-PUB-161116-2 PMID:27845735

Deletion of Pr130 Interrupts Cardiac Development in Zebrafish

Yang et al., 2016

ZDB-PUB-161116-2
PMID:27845735