PUBLICATION

Distinct roles for the cell adhesion molecule Contactin2 in the development and function of neural circuits in zebrafish

Authors

Gurung, S., Asante, E., Hummel, D., Williams, A., Feldman-Schultz, O., Halloran, M.C., Sittaramane, V., Chandrasekhar, A.

ID

ZDB-PUB-180520-6

Date

2018

Source

Mechanisms of Development 152: 1-12 (Journal)

Registered Authors

Chandrasekhar, Anand, Halloran, Mary, Sittaramane, Vinoth

Keywords

Axon guidance, CRISPR/Cas9, Cell adhesion molecule, Cntn2, Facial branchiomotor neuron, Morpholino, Neuronal migration, Vangl2, Zebrafish, nucMLF

MeSH Terms

Zebrafish/genetics*
Zebrafish/growth & development
Contactin 2/genetics*
Cell Adhesion/genetics*
Humans
Mice
Motor Neurons/metabolism
Cell Polarity/genetics
Gene Expression Regulation, Developmental/genetics
CRISPR-Cas Systems
Morpholinos/genetics
Morpholinos/metabolism
Cell Movement/genetics
Animals
Axons/metabolism
Neurogenesis/genetics*
Zebrafish Proteins/genetics*

(all 17)

PubMed

29777776 Full text @ Mech. Dev.

Abstract

Contactin2 (Cntn2)/Transient Axonal Glycoprotein 1 (Tag1), a neural cell adhesion molecule, has established roles in neuronal migration and axon fasciculation in chick and mouse. In zebrafish, antisense morpholino-based studies have indicated roles for cntn2 in the migration of facial branchiomotor (FBM) neurons, the guidance of the axons of the nucleus of the medial longitudinal fascicle (nucMLF), and the outgrowth of Rohon-Beard (RB) central axons. To study functions of Cntn2 in later stages of neuronal development, we generated cntn2 mutant zebrafish using CRISPR-Cas9. Using a null mutant allele, we detected genetic interactions between cntn2 and the planar cell polarity gene vangl2, as shown previously with cntn2 morphants, demonstrating a function for cntn2 during FBM neuron migration in a sensitized background of reduced planar cell polarity signaling. In addition, maternal-zygotic (MZ) cntn2 mutant larvae exhibited aberrant touch responses and swimming, suggestive of defects in sensorimotor circuits, consistent with studies in mice. However, the nucMLF axon convergence FBM neuron migration, and RB outgrowth defects seen in morphants were not seen in the mutants, and we show here that they are likely off-target effects of morpholinos. However, MLF axons exhibited local defasciculation in MZcntn2 mutants, consistent with a role for Cntn2 in axon fasciculation. These data demonstrate distinct roles for zebrafish cntn2 in neuronal migration and axon fasciculation, and in the function of sensorimotor circuits.

Genes / Markers

Figures

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Expression

Gene	Antibody	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
cntn2		cntn2^zou20/zou20	standard conditions	Prim-15		anterior lateral line ganglion	ISH	Fig. 1
cntn2	ab1-cntn2	cntn2^zou20/zou20	standard conditions	Prim-5	Not Detected	anterior lateral line ganglion	IHC	Fig. 1
cntn2	ab1-cntn2	cntn2^zou20/zou20	standard conditions	Prim-5	Not Detected	posterior lateral line ganglion	IHC	Fig. 1
cntn2		cntn2^zou20/zou20	standard conditions	Prim-15		posterior lateral line ganglion	ISH	Fig. 1
cntn2	ab1-cntn2	cntn2^zou20/zou20	standard conditions	Prim-5	Not Detected	statoacoustic (VIII) ganglion	IHC	Fig. 1
cntn2		cntn2^zou20/zou20	standard conditions	Prim-15		statoacoustic (VIII) ganglion	ISH	Fig. 1
cntn2	ab1-cntn2	cntn2^zou20/zou20	standard conditions	Long-pec	Not Detected	whole organism	WB	Fig. 1
cntn2	ab1-cntn2	cntn2^zou22/zou22	standard conditions	Long-pec	Not Detected	whole organism	WB	Fig. 1
cntn2		WT	standard conditions	Prim-15		anterior lateral line ganglion	ISH	Fig. 1
cntn2	ab1-cntn2	WT	standard conditions	Prim-5		anterior lateral line ganglion	IHC	Fig. 1

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
cntn2^zou20/+	standard conditions	Days 7-13	swimming duration, normal	Fig. 7
cntn2^zou20/zou20	standard conditions	Prim-5	anterior lateral line ganglion cntn2 expression absent, abnormal	Fig. 1
cntn2^zou20/zou20	standard conditions	Prim-5	medial longitudinal fasciculus axonal fasciculation decreased occurrence, abnormal	Fig. 4
cntn2^zou20/zou20	standard conditions	Prim-5	nucleus of the medial longitudinal fasciculus medulla oblongata neuron aggregated, normal	Fig. 4
cntn2^zou20/zou20	standard conditions	Prim-5	posterior lateral line ganglion cntn2 expression absent, abnormal	Fig. 1
cntn2^zou20/zou20	standard conditions	Prim-5	statoacoustic (VIII) ganglion cntn2 expression absent, abnormal	Fig. 1
cntn2^zou20/zou20	standard conditions	Prim-15	anterior lateral line ganglion cntn2 expression decreased amount, abnormal	Fig. 1
cntn2^zou20/zou20	standard conditions	Prim-15	posterior lateral line ganglion cntn2 expression decreased amount, abnormal	Fig. 1
cntn2^zou20/zou20	standard conditions	Prim-15	statoacoustic (VIII) ganglion cntn2 expression decreased amount, abnormal	Fig. 1
cntn2^zou20/zou20	standard conditions	Long-pec	head startle response occurrence, normal	Fig. 6

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
rw0Tg	Tg(isl1a:GFP)	Transgenic Insertion
tc240		Insertion	vangl2
ti263a		Unknown	lamc1
zou20		Indel	cntn2
zou22		Small Deletion	cntn2
zy8Tg	Tg(pitx2:EGFP)	Transgenic Insertion

1 - 6 of 6

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
cntn2	CRISPR2-cntn2	CRISPR
cntn2	MO1-cntn2	MRPHLNO

Fish

Fish
cntn2^zou20/+
cntn2^zou20/+
cntn2^zou20/+; rw0Tg
cntn2^zou20/+; rw0Tg
cntn2^zou20/zou20
cntn2^zou20/zou20
cntn2^zou20/zou20; rw0Tg
cntn2^zou20/zou20; rw0Tg
cntn2^zou20/zou20; rw0Tg + MO1-cntn2
cntn2^zou20/zou20; zy8Tg

1 - 10 of 24

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GFP	EFG	GFP

Mapping

Gurung et al., 2018 ZDB-PUB-180520-6 PMID:29777776

Distinct roles for the cell adhesion molecule Contactin2 in the development and function of neural circuits in zebrafish

Gurung et al., 2018

ZDB-PUB-180520-6
PMID:29777776