PUBLICATION

Graded effects of unregulated smooth muscle myosin on intestinal architecture, intestinal motility, and vascular function in zebrafish

Authors

Abrams, J., Einhorn, Z., Seiler, C., Zong, A.B., Sweeney, H.L., Pack, M.

ID

ZDB-PUB-160220-3

Date

2016

Source

Disease models & mechanisms 9(5): 529-40 (Journal)

Registered Authors

Abrams, Joshua, Einhorn, Zev, Pack, Michael, Seiler, Christoph

Keywords

Myosin, Zebrafish, Intestine, Smooth muscle

MeSH Terms

Genetic Testing
Neovascularization, Physiologic*
Mutation/genetics
Amino Acid Sequence
Alleles
Zebrafish/metabolism*
Gastrointestinal Motility*
Heterozygote
Homozygote
Genes, Dominant
Intestines/anatomy & histology*
Intestines/blood supply*
Intestines/physiology
Sequence Analysis, DNA
Exome/genetics
Animals
Oxidation-Reduction
Myosin Heavy Chains/chemistry
Myosin Heavy Chains/genetics
Myosin Heavy Chains/metabolism*
Oxidative Stress
Zebrafish Proteins/chemistry
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*

(all 24)

PubMed

26893369 Full text @ Dis. Model. Mech.

Abstract

Smooth muscle contraction is controlled by the regulated activity of the myosin heavy chain (Myh11) ATPase. Myh11 mutations have diverse effects in the cardiovascular, digestive and genitourinary systems in humans and animal models. We previously reported a recessive missense mutation, meltdown (mlt) that converts a highly conserved tryptophan to arginine (W512R) in the rigid relay loop of zebrafish Myh11. The mlt mutation disrupts myosin regulation and non-autonomously induces invasive expansion of the intestinal epithelium. Here we report two novel missense mutations in the Switch-1 (S237Y) and coil-coiled (L1287M) domains of Myh11 that fail to complement mlt. Cell invasion was not detected in either homozygous mutant but could be induced by oxidative stress and activation of oncogenic signaling pathways. The smooth muscle defect imparted by the mlt and S237Y mutations also delayed intestinal transit, and altered vascular function, as measured by blood flow in the dorsal aorta. The cell invasion phenotype induced by the three myh11 mutants correlated with the degree of myosin deregulation. These findings suggest that the vertebrate intestinal epithelium is tuned to the physical state of the surrounding stroma, which in turn, governs its response to physiologic and pathologic stimuli. Genetic variants that alter regulation of smooth muscle myosin may be risk factors for diseases affecting the intestine, vasculature and other tissues that contain smooth muscle or contractile cells that express smooth muscle proteins, particularly in the setting of redox stress.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
GFP	myh11a^m498/+; myh11a^p318/+; p306Tg	standard conditions	Protruding-mouth	intestinal epithelium actin-based cell projection	IHC	Fig. 2
GFP	myh11a^m498/+; myh11a^p318/+; p306Tg	standard conditions	Day 4	intestinal epithelium actin-based cell projection	IHC	Fig. 2
GFP	myh11a^m498/+; myh11a^p318/+; p306Tg	standard conditions	Protruding-mouth	intestinal epithelium	IHC	Fig. 2
GFP	myh11a^m498/+; myh11a^p318/+; p306Tg	standard conditions	Day 4	intestinal epithelium	IHC	Fig. 2
GFP	p306Tg	standard conditions	Protruding-mouth	intestinal epithelium	IHC	Fig. 2
GFP	p306Tg	standard conditions	Day 4	intestinal epithelium	IHC	Fig. 2

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment: menadione	Protruding-mouth	intestine morphology, normal	Fig. 5
WT + MO1-acta2	standard conditions	Day 4	artery increased diameter, abnormal	Fig. 7
WT + MO1-acta2	standard conditions	Day 4	blood circulation increased volume, abnormal	Fig. 7
WT + MO1-acta2	standard conditions	Day 4	heart contraction increased frequency, abnormal	Fig. 7
WT + MO1-acta2	standard conditions	Day 4	vasodilation increased occurrence, abnormal	Fig. 7
myh11a^m498/+	standard conditions	Day 4	artery decreased diameter, abnormal	Fig. 7
myh11a^m498/+	standard conditions	Day 4	blood circulation decreased volume, abnormal	Fig. 7
myh11a^m498/+	standard conditions	Day 4	heart contraction increased frequency, abnormal	Fig. 7
myh11a^m498/+	standard conditions	Day 4	vasoconstriction increased occurrence, abnormal	Fig. 7
myh11a^m498/+	chemical treatment: menadione	Protruding-mouth	mid intestine epithelial cell migration increased occurrence, abnormal	Fig. 5

1 - 10 of 52

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
m498		Point Mutation	myh11a
p151Tg	Tg(tagln:GFP)	Transgenic Insertion
p306Tg	Tg(mir194:LIFEACT-GFP)	Transgenic Insertion
p309Tg	Tg(mir194:mCherry)	Transgenic Insertion
p311Tg	Tg(mir194:GFP-Hsa.KRAS_G12V)	Transgenic Insertion
p318		Point Mutation	myh11a
p319		Point Mutation	myh11a
tm213		Point Mutation	axin1

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Human Disease / Model

Human Disease	Fish	Conditions	Evidence
megacystis-microcolon-intestinal hypoperistalsis syndrome	myh11a^m498/+; myh11a^p319/+	standard conditions	TAS
megacystis-microcolon-intestinal hypoperistalsis syndrome	myh11a^m498/+; myh11a^p318/+	standard conditions	TAS
obsolete Batten Turner congenital myopathy			TAS

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Sequence Targeting Reagents

Target	Reagent	Reagent Type
acta2	MO1-acta2	MRPHLNO
cald1a	MO1-cald1a	MRPHLNO
myh11a	MO1-myh11a	MRPHLNO

Fish

Fish
axin1^tm213/+; myh11a^p319/+; p311Tg
axin1^tm213/tm213; myh11a^m498/+; p311Tg
axin1^tm213/tm213; p311Tg
myh11a^m498/m498
myh11a^m498/m498 + MO1-myh11a
myh11a^m498/+
myh11a^m498/+; myh11a^p318/+
myh11a^m498/+; myh11a^p318/+; p306Tg
myh11a^m498/+; myh11a^p319/+
myh11a^p318/p318

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Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab1-lama1	polyclonal			Rabbit
Ab2-krt	monoclonal		IgG1	Mouse

Orthology

Gene	Orthology
myh11a

Engineered Foreign Genes

Marker	Marker Type	Name
GFP	EFG	GFP
mCherry	EFG	mCherry

Mapping

Abrams et al., 2016 ZDB-PUB-160220-3 PMID:26893369

Graded effects of unregulated smooth muscle myosin on intestinal architecture, intestinal motility, and vascular function in zebrafish

Abrams et al., 2016

ZDB-PUB-160220-3
PMID:26893369