PUBLICATION

CaMK-II activation is essential for zebrafish inner ear development and acts through Delta-Notch signaling

Authors

Rothschild, S.C., Lahvic, J., Francescatto, L., McLeod, J.J., Burgess, S.M., and Tombes, R.M.

ID

ZDB-PUB-130709-33

Date

2013

Source

Developmental Biology 381(1): 179-88 (Journal)

Registered Authors

Burgess, Shawn, Francescatto, Ludmila, McLeod, Jamie, Rothschild, Sarah Chase, Tombes, Robert M.

Keywords

CaMK-II, cilia, otolith, inner ear, Delta-notch

MeSH Terms

Animals
Phosphorylation
Oligonucleotides/metabolism
Enzyme Activation
Intracellular Signaling Peptides and Proteins/metabolism*
Membrane Proteins/metabolism*
Zebrafish/embryology*
Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism*
Exocytosis
Gene Expression Regulation, Developmental*
Receptors, Notch/metabolism*
Ear, Inner/embryology*
Cilia/metabolism
Signal Transduction

(all 14)

PubMed

23747599 Full text @ Dev. Biol.

Abstract

Zebrafish inner ear development is characterized by the crystallization of otoliths onto immotile kinocilia that protrude from sensory “hair” cells. The stereotypical formation of these sensory structures is dependent on the expression of key patterning genes and on Ca²⁺ signals. One potential target of Ca²⁺ signaling in the inner ear is the type II Ca²⁺/calmodulin-dependent protein kinase (CaMK-II), which is preferentially activated in hair cells, with intense activation at the base of kinocilia. In zebrafish, CaMK-II is encoded by seven genes; the expression of one of these genes (camk2g1) is enriched in hair cells. The suppression of camk2g1 expression by antisense morpholino oligonucleotides or inhibition of CaMK-II activation by the pharmacological antagonist, KN-93, results in aberrant otolith formation without preventing cilia formation. In fact, CaMK-II suppression results in additional ciliated hair cells and altered levels of Delta–Notch signaling members. DeltaA and deltaD transcripts are increased and DeltaD protein accumulates in hair cells of CaMK-II morphants, indicative of defective recycling and/or exocytosis. Our findings indicate that CaMK-II plays a critical role in the developing ear, influencing cell differentiation through extranuclear effects on Delta–Notch signaling. Continued expression and activation of CaMK-II in maculae and cristae in older embryos suggests continued roles in auditory sensory maturation and transduction.

Genes / Markers

Figures

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Expression

Gene	Antibody	Fish	Conditions	Stage	Anatomy	Assay	Figure
dla		WT	control	Prim-5	otic vesicle	ISH	Fig. 6
dla		WT + MO1-camk2g1	standard conditions	Prim-5	otic vesicle	ISH	Fig. 6
dld	ab1-dld	WT	control	Prim-5	auditory receptor cell	IHC	Fig. 6
dld		WT	control	Prim-5	otic vesicle	ISH	Fig. 6
dld	ab1-dld	WT + MO1-camk2g1	standard conditions	Prim-5	auditory receptor cell	IHC	Fig. 6
dld		WT + MO1-camk2g1	standard conditions	Prim-5	otic vesicle	ISH	Fig. 6
gata3		WT	chemical treatment: pharmaceutical	Prim-5	otic vesicle	ISH	Fig. 3
gata3		WT	control	Prim-5	otic vesicle	ISH	Fig. 3
GFP		vu119Tg	standard conditions	Prim-5	auditory receptor cell	IFL	Fig. 1
pax2a		WT	control	Prim-5	otic vesicle	ISH	Fig. 6

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment: pharmaceutical	Prim-15	otolith decreased size, abnormal	Fig. 3
WT	chemical treatment: pharmaceutical	Prim-15	otolith mislocalised, abnormal	Fig. 3
WT	chemical treatment: pharmaceutical	Prim-15	otolith morphology, abnormal	Fig. 3
WT	chemical treatment: pharmaceutical	Protruding-mouth	otolith decreased size, abnormal	Fig. 3
WT	chemical treatment: pharmaceutical	Protruding-mouth	otolith mislocalised, abnormal	Fig. 3
WT	chemical treatment: pharmaceutical	Protruding-mouth	otolith morphology, abnormal	Fig. 3
WT + MO1-camk2g1	standard conditions	Prim-5	otic vesicle kinocilium increased amount, abnormal	Fig. 4
WT + MO1-camk2g1	standard conditions	Prim-5	otic vesicle kinocilium increased amount, abnormal	Fig. 6
WT + MO1-camk2g1	standard conditions	Prim-15	otolith decreased size, abnormal	Fig. 2
WT + MO1-camk2g1	standard conditions	Prim-15	otolith mislocalised, abnormal	Fig. 2

1 - 10 of 19

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
vu119Tg	Tg(Ola.Actb:Hsa.HRAS-EGFP)	Transgenic Insertion

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
camk2g1	MO1-camk2g1	MRPHLNO

Fish

Fish
vu119Tg
WT
WT + MO1-camk2g1

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
GFP	EFG	GFP

Mapping

Rothschild et al., 2013 ZDB-PUB-130709-33 PMID:23747599

CaMK-II activation is essential for zebrafish inner ear development and acts through Delta-Notch signaling

Rothschild et al., 2013

ZDB-PUB-130709-33
PMID:23747599