PUBLICATION

Two knockdown models of the autism genes SYNGAP1 and SHANK3 in zebrafish produce similar behavioral phenotypes associated with embryonic disruptions of brain morphogenesis

Authors

Kozol, R.A., Cukier, H.N., Zou, B., Mayo, V., De Rubeis, S., Cai, G., Griswold, A.J., Whitehead, P.L., Haines, J.L., Gilbert, J.R., Cuccaro, M.L., Martin, E.R., Baker, J.D., Buxbaum, J.D., Pericak-Vance, M.A., Dallman, J.E.

ID

ZDB-PUB-150418-11

Date

2015

Source

Human molecular genetics 24(14): 4006-23 (Journal)

Registered Authors

Dallman, Julia

Keywords

none

MeSH Terms

Animals
Autism Spectrum Disorder/genetics*
Brain/embryology*
Databases, Genetic
Embryonic Development
GTPase-Activating Proteins/genetics*
GTPase-Activating Proteins/metabolism
Gene Duplication
Gene Expression Regulation, Developmental
Gene Knockdown Techniques
Haploinsufficiency
Nerve Tissue Proteins/genetics*
Nerve Tissue Proteins/metabolism
Organogenesis/genetics*
Phenotype
Zebrafish/embryology
Zebrafish/genetics*
Zebrafish Proteins/genetics*
Zebrafish Proteins/metabolism
ras GTPase-Activating Proteins/genetics*
ras GTPase-Activating Proteins/metabolism

(all 21)

PubMed

25882707 Full text @ Hum. Mol. Genet.

Abstract

Despite significant progress in the genetics of autism spectrum disorder (ASD), how genetic mutations translate to the behavioral changes characteristic of ASD remains largely unknown. ASD affects 1-2% of children and adults, and is characterized by deficits in verbal and non-verbal communication, and social interactions, as well as the presence of repetitive behaviors and/or stereotyped interests. ASD is clinically and etiologically heterogeneous, with a strong genetic component. Here, we present functional data from syngap1 and shank3 zebrafish loss-of-function models of ASD. SYNGAP1, a synaptic Ras GTPase activating protein, and SHANK3, a synaptic scaffolding protein, were chosen because of mounting evidence that haploinsufficiency in these genes is highly penetrant for ASD and intellectual disability (ID). Orthologs of both SYNGAP1 and SHANK3 are duplicated in the zebrafish genome and we find that all four transcripts (syngap1a, syngap1b, shank3a and shank3b) are expressed at the earliest stages of nervous system development with pronounced expression in the larval brain. Consistent with early expression of these genes, knockdown of syngap1b or shank3a cause common embryonic phenotypes including delayed mid- and hindbrain development, disruptions in motor behaviors that manifest as unproductive swim attempts, and spontaneous, seizure-like behaviors. Our findings indicate that both syngap1b and shank3a play novel roles in morphogenesis resulting in common brain and behavioral phenotypes.

Genes / Markers

Marker	Marker Type	Name
shank3a	GENE	SH3 and multiple ankyrin repeat domains 3a
shank3b	GENE	SH3 and multiple ankyrin repeat domains 3b
syngap1a	GENE	synaptic Ras GTPase activating protein 1a
syngap1b	GENE	synaptic Ras GTPase activating protein 1b

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Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
shank3a	WT	standard conditions	Long-pec	forebrain	ISH	Fig. 2
shank3a	WT	standard conditions	Long-pec	hindbrain	ISH	Fig. 2
shank3a	WT	standard conditions	Long-pec	midbrain	ISH	Fig. 2
shank3a	WT	standard conditions	64-cell to Day 5	whole organism	RTPCR	Fig. 2
shank3b	WT	standard conditions	Long-pec	forebrain	ISH	Fig. 2
shank3b	WT	standard conditions	Long-pec	midbrain	ISH	Fig. 2
shank3b	WT	standard conditions	64-cell to Day 5	whole organism	RTPCR	Fig. 2
syngap1a	WT	standard conditions	Long-pec	forebrain anterior region	ISH	Fig. 2
syngap1a	WT	standard conditions	Long-pec	hindbrain dorsal region	ISH	Fig. 2
syngap1a	WT	standard conditions	64-cell to Day 5	whole organism	RTPCR	Fig. 2

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT + MO1-syngap1a + MO1-syngap1b	standard conditions	Long-pec	swimming behavior disrupted, abnormal	Fig. 3
WT + MO1-syngap1a + MO1-syngap1b	standard conditions	Protruding-mouth	swimming behavior disrupted, abnormal	Fig. 4
WT + MO1-syngap1a + MO1-syngap1b	standard conditions	Protruding-mouth	thigmotaxis disrupted, abnormal	Fig. 4
WT + MO1-syngap1b	standard conditions	Prim-5	midbrain hindbrain boundary morphology, abnormal	Fig. 5
WT + MO1-syngap1b	standard conditions	Long-pec	brain decreased size, abnormal	Fig. 6
WT + MO1-syngap1b	standard conditions	Long-pec	brain development delayed, abnormal	Fig. 5
WT + MO1-syngap1b	standard conditions	Long-pec	embryo development delayed, abnormal	Fig. 5
WT + MO1-syngap1b	standard conditions	Long-pec	hindbrain GABAergic neuron decreased amount, abnormal	Fig. 6
WT + MO1-syngap1b	standard conditions	Long-pec	hindbrain glutamatergic neuron decreased amount, abnormal	Fig. 6
WT + MO1-syngap1b	standard conditions	Long-pec	midbrain GABAergic neuron decreased amount, abnormal	Fig. 6

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
nksaigff213aGt	Gt(IRES-GAL4FF)	Transgenic Insertion
nns14Tg	TgBAC(slc17a6b:LOXP-DsRed-LOXP-GFP)	Transgenic Insertion

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
shank3a	MO2-shank3a	MRPHLNO
shank3b	MO3-shank3b	MRPHLNO
syngap1a	MO1-syngap1a	MRPHLNO
syngap1b	MO1-syngap1b	MRPHLNO

Fish

Fish
nksaigff213aGt; nns14Tg + MO1-syngap1b
nksaigff213aGt; nns14Tg + MO1-syngap1b + MO2-shank3a
nksaigff213aGt; nns14Tg + MO2-shank3a
WT
WT + MO1-syngap1a + MO1-syngap1b
WT + MO1-syngap1b
WT + MO1-syngap1b + MO2-shank3a
WT + MO2-shank3a

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Antibodies

Orthology

Gene	Orthology
shank3a
shank3b
syngap1a
syngap1b

Engineered Foreign Genes

Marker	Marker Type	Name
DsRed	EFG	DsRed
GAL4FF	EFG	GAL4FF
GFP	EFG	GFP

Mapping

Kozol et al., 2015 ZDB-PUB-150418-11 PMID:25882707

Two knockdown models of the autism genes SYNGAP1 and SHANK3 in zebrafish produce similar behavioral phenotypes associated with embryonic disruptions of brain morphogenesis

Kozol et al., 2015

ZDB-PUB-150418-11
PMID:25882707