PUBLICATION

Loss of Sec-1 Family Domain-Containing 1 (scfd1) Causes Severe Cardiac Defects and Endoplasmic Reticulum Stress in Zebrafish

Authors: Huttner, I.G., Santiago, C.F., Jacoby, A., Cheng, D., Trivedi, G., Cull, S., Cvetkovska, J., Chand, R., Berger, J., Currie, P.D., Smith, K.A., Fatkin, D.
ID: ZDB-PUB-231028-53
Date: 2023
Source: Journal of cardiovascular development and disease 10(10): (Journal)
Registered Authors: Berger, Joachim, Chand, Renee, Currie, Peter D., Fatkin, Diane, Huttner, Inken, Trivedi, Gunjan
Keywords: ER stress, dilated cardiomyopathy, genetics, protein homeostasis, scfd1, zebrafish
MeSH Terms: none
PubMed: 37887855 Full text @ J Cardiovasc Dev Dis

Abstract

Dilated cardiomyopathy (DCM) is a common heart muscle disorder that frequently leads to heart failure, arrhythmias, and death. While DCM is often heritable, disease-causing mutations are identified in only ~30% of cases. In a forward genetic mutagenesis screen, we identified a novel zebrafish mutant, heart and head (hah^vcc43), characterized by early-onset cardiomyopathy and craniofacial defects. Linkage analysis and next-generation sequencing identified a nonsense variant in the highly conserved scfd1 gene, also known as sly1, that encodes sec1 family domain-containing 1. Sec1/Munc18 proteins, such as Scfd1, are involved in membrane fusion regulating endoplasmic reticulum (ER)/Golgi transport. CRISPR/Cas9-engineered scfd1^vcc44 null mutants showed severe cardiac and craniofacial defects and embryonic lethality that recapitulated the phenotype of hah^vcc43 mutants. Electron micrographs of scfd1-depleted cardiomyocytes showed reduced myofibril width and sarcomere density, as well as reticular network disorganization and fragmentation of Golgi stacks. Furthermore, quantitative PCR analysis showed upregulation of ER stress response and apoptosis markers. Both heterozygous hah^vcc43 mutants and scfd1^vcc44 mutants survived to adulthood, showing chamber dilation and reduced ventricular contraction. Collectively, our data implicate scfd1 loss-of-function as the genetic defect at the hah^vcc43 locus and provide new insights into the role of scfd1 in cardiac development and function.

Genes / Markers

Figures

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Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + MO1-scfd1	control	Protruding-mouth	embryonic heart tube formation normal process quality, normal	Figure 2
WT + MO1-scfd1	control	Protruding-mouth	eye decreased size, abnormal	Figure 2
WT + MO1-scfd1	control	Protruding-mouth	heart looping normal process quality, normal	Figure 2
WT + MO1-scfd1	control	Protruding-mouth	heart morphology, abnormal	Figure 2
WT + MO1-scfd1	control	Protruding-mouth	pericardium edematous, abnormal	Figure 2
WT + MO1-scfd1	control	Protruding-mouth	ventral mandibular arch absence of anatomical entity, abnormal	Figure 2
scfd1^vcc43/+	standard conditions	Protruding-mouth	whole organism Ab1-scfd1 labeling decreased amount, abnormal	Figure 2
scfd1^vcc43/+	standard conditions	Day 4	heart contraction normal process quality, normal	Figure 3
scfd1^vcc43/+	standard conditions	Day 4	heart contraction rate of continuous process, normal	Figure 3
scfd1^vcc43/+	standard conditions	Day 4	whole organism viable, normal	Figure 3

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
vcc43		Point Mutation	scfd1
vcc44		Small Deletion	scfd1

Human Disease / Model

Human Disease	Fish	Conditions	Evidence
dilated cardiomyopathy			TAS

Sequence Targeting Reagents

Target	Reagent	Reagent Type
scfd1	CRISPR1-scfd1	CRISPR
scfd1	MO1-scfd1	MRPHLNO
tyr	CRISPR54-tyr	CRISPR

Fish

Fish
scfd1^vcc43/+
scfd1^vcc43/vcc43
scfd1^vcc44/+
scfd1^vcc44/vcc44
WT
WT + MO1-scfd1

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Huttner et al., 2023 ZDB-PUB-231028-53 PMID:37887855

Loss of Sec-1 Family Domain-Containing 1 (scfd1) Causes Severe Cardiac Defects and Endoplasmic Reticulum Stress in Zebrafish

Huttner et al., 2023

ZDB-PUB-231028-53
PMID:37887855