PUBLICATION

Knockout of zebrafish desmin genes does not cause skeletal muscle degeneration but alters calcium flux

Authors

Kayman Kürekçi, G., Kural Mangit, E., Koyunlar, C., Unsal, S., Saglam, B., Ergin, B., Gizer, M., Uyanik, I., Boustanabadimaralan Düz, N., Korkusuz, P., Talim, B., Purali, N., Hughes, S.M., Dincer, P.R.

ID

ZDB-PUB-210407-73

Date

2021

Source

Scientific Reports 11: 7505 (Journal)

Registered Authors

Hughes, Simon M.

Keywords

none

MeSH Terms

Gene Expression Profiling
Animals
Desmin/genetics*
Desmin/metabolism
Mutation/genetics
Base Sequence
RNA, Messenger/genetics
RNA, Messenger/metabolism
Neuromuscular Junction/pathology
Gene Expression Regulation, Developmental
Zebrafish/embryology
Zebrafish/genetics*
Calcium/metabolism*
Embryo, Nonmammalian/metabolism
Muscle Fibers, Skeletal/pathology
Muscle, Skeletal/metabolism*
Muscle, Skeletal/pathology*
Muscle, Skeletal/ultrastructure
Gene Knockout Techniques*
Larva/genetics

(all 20)

PubMed

33820917 Full text @ Sci. Rep.

Abstract

Desmin is a muscle-specific intermediate filament protein that has fundamental role in muscle structure and force transmission. Whereas human desmin protein is encoded by a single gene, two desmin paralogs (desma and desmb) exist in zebrafish. Desma and desmb show differential spatiotemporal expression during zebrafish embryonic and larval development, being similarly expressed in skeletal muscle until hatching, after which expression of desmb shifts to gut smooth muscle. We generated knockout (KO) mutant lines carrying loss-of-function mutations for each gene by using CRISPR/Cas9. Mutants are viable and fertile, and lack obvious skeletal muscle, heart or intestinal defects. In contrast to morphants, knockout of each gene did not cause any overt muscular phenotype, but did alter calcium flux in myofibres. These results point to a possible compensation mechanism in these mutant lines generated by targeting nonsense mutations to the first coding exon.

Genes / Markers

Figures

Show all Figures

Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
desma^kg97/+ (AB)	standard conditions	Long-pec	thigmotaxis normal process quality, normal	Figure 4
desma^kg97/kg97 (AB)	standard conditions	1-cell to Day 5	whole organism viable, normal	Figure 4
desma^kg97/kg97 (AB)	standard conditions	Long-pec	hatching normal rate, normal	Figure 4
desma^kg97/kg97 (AB)	standard conditions	Long-pec	thigmotaxis normal process quality, normal	Figure 4
desma^kg97/kg97 (AB)	standard conditions	Protruding-mouth	hatching normal rate, normal	Figure 4
desma^kg97/kg97 (AB)	standard conditions	Day 4	myotome ab1-desm labeling decreased amount, abnormal	Figure 3
desma^kg97/kg97 (AB)	standard conditions	Day 4	myotome ab1-desm labeling decreased distribution, abnormal	Figure 3
desma^kg97/kg97 (AB)	standard conditions	Day 4	whole organism desma expression decreased amount, abnormal	Figure 2
desma^kg97/kg97 (AB)	standard conditions	Day 4	whole organism desmb expression increased amount, abnormal	Figure 2
desma^kg97/kg97 (AB)	standard conditions	Day 4	whole organism morphology, normal	Figure 2

1 - 10 of 36

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
kg97		Indel	desma
kg156		Small Deletion	desmb

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
desma	CRISPR1-desma	CRISPR
desmb	CRISPR1-desmb	CRISPR

Fish

Fish
AB
desma^kg97/kg97 (AB)
desma^kg97/+ (AB)
desmb^kg156/kg156 (AB)
desmb^kg156/kg156; desma^kg97/kg97 (AB)
desmb^kg156/kg156; desma^kg97/+ (AB)
desmb^kg156/+ (AB)

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-desm	polyclonal	desma		Rabbit
Ab5-lmnb2	polyclonal		IgG	Chicken

Orthology

Engineered Foreign Genes

Mapping

Kayman Kürekçi et al., 2021 ZDB-PUB-210407-73 PMID:33820917

Knockout of zebrafish desmin genes does not cause skeletal muscle degeneration but alters calcium flux

Kayman Kürekçi et al., 2021

ZDB-PUB-210407-73
PMID:33820917