PUBLICATION

Stathmin-like 4 is critical for the maintenance of neural progenitor cells in dorsal midbrain of zebrafish larvae

Authors

Lin, M.J., Lee, S.J.

ID

ZDB-PUB-161108-3

Date

2016

Source

Scientific Reports 6: 36188 (Journal)

Registered Authors

Lee, Shyh-Jye

Keywords

Developmental neurogenesis, Neural progenitors

MeSH Terms

Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Zebrafish/embryology*
Zebrafish/genetics
Cell Differentiation*
Stathmin/genetics
Stathmin/metabolism*
Mesencephalon/cytology
Mesencephalon/embryology*
Neural Stem Cells/cytology
Neural Stem Cells/metabolism*
Animals
Wnt Signaling Pathway*

(all 13)

PubMed

27819330 Full text @ Sci. Rep.

Abstract

A delicate balance between proliferating and differentiating signals is necessary to ensure proper growth and neuronal specification. By studying the developing zebrafish brain, we observed a specific and dynamic expression of a microtubule destabilizer gene, stathmin-like 4 (stmn4), in the dorsal midbrain region. The expression of stmn4 was mutually exclusive to a pan-neuronal marker, elavl3 that indicates its role in regulating neurogenesis. We showed the knockdown or overexpression of stmn4 resulted in premature neuronal differentiation in dorsal midbrain. We also generated stmn4 maternal-zygotic knockout zebrafish by the CRISPR/Cas9 system. Unexpectedly, only less than 10% of stmn4 mutants showed similar phenotypes observed in that of stmn4 morphants. It might be due to the complementation of the increased stmn1b expression observed in stmn4 mutants. In addition, time-lapse recordings revealed the changes in cellular proliferation and differentiation in stmn4 morphants. Stmn4 morphants displayed a longer G₂ phase that could be rescued by Cdc25a. Furthermore, the inhibition of Wnt could reduce stmn4 transcripts. These results suggest that the Wnt-mediated Stmn4 homeostasis is crucial for preventing dorsal midbrain from premature differentiation via the G₂ phase control during the neural keel stage.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
elavl3	AB	standard conditions	14-19 somites	trigeminal sensory neuron	ISH	Fig. 1
elavl3	AB	standard conditions	10-13 somites	trigeminal sensory neuron	ISH	Fig. 1
elavl3	AB + MO1-stmn4	standard conditions	10-13 somites	midbrain dorsal region	ISH	Fig. 2
elavl3	AB + MO2-stmn4	standard conditions	10-13 somites	midbrain dorsal region	ISH	Fig. 2
elavl3	stmn4^{twu0101/twu0101}; rw0130aTg + MO2-stmn4	standard conditions	14-19 somites	midbrain dorsal region	ISH	Fig. 3
elavl3	stmn4^{twu0102/twu0102}; rw0130aTg + MO2-stmn4	standard conditions	14-19 somites	midbrain dorsal region	ISH	Fig. 3
Kaede	rw0130aTg	standard conditions	14-19 somites	epiphysis neuron	IFL	Fig. 2
Kaede	rw0130aTg	standard conditions	14-19 somites	trigeminal sensory neuron	IFL	Fig. 2
Kaede	rw0130aTg + MO1-stmn4	standard conditions	14-19 somites	epiphysis neuron	IFL	Fig. 2
Kaede	rw0130aTg + MO1-stmn4	standard conditions	14-19 somites	midbrain dorsal region	IFL	Fig. 2

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + MO1-stmn4	standard conditions	10-13 somites	midbrain dorsal region elavl3 expression increased amount, abnormal	Fig. 2
AB + MO1-stmn4	standard conditions	10-13 somites	midbrain dorsal region elavl3 expression mislocalised, abnormal	Fig. 2
AB + MO2-stmn4	standard conditions	10-13 somites	midbrain dorsal region elavl3 expression increased amount, abnormal	Fig. 2
AB + MO2-stmn4	standard conditions	10-13 somites	midbrain dorsal region elavl3 expression mislocalised, abnormal	Fig. 2
AB + MO2-stmn4	standard conditions	14-19 somites	cell population proliferation decreased occurrence, abnormal	Fig. 6
AB + MO2-stmn4	standard conditions	14-19 somites	mitotic G2 phase increased duration, abnormal	Fig. 6
rw0130aTg + MO1-stmn4	standard conditions	14-19 somites	midbrain dorsal region Kaede expression increased amount, abnormal	Fig. 2
rw0130aTg + MO1-stmn4	standard conditions	14-19 somites	midbrain dorsal region Kaede expression mislocalised, abnormal	Fig. 2
rw0130aTg + MO2-stmn4	standard conditions	14-19 somites	midbrain dorsal region Kaede expression increased amount, abnormal	Fig. 2
rw0130aTg + MO2-stmn4	standard conditions	14-19 somites	midbrain dorsal region Kaede expression mislocalised, abnormal	Fig. 2

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
rw0130aTg	Tg(elavl3:Kaede)	Transgenic Insertion
twu0101		Small Deletion	stmn4
twu0102		Small Deletion	stmn4
w32Tg	Tg(hsp70l:dkk1b-GFP)	Transgenic Insertion

1 - 4 of 4

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
stmn4	CRISPR1-stmn4	CRISPR
stmn4	CRISPR2-stmn4	CRISPR
stmn4	CRISPR3-stmn4	CRISPR
stmn4	MO1-stmn4	MRPHLNO
stmn4	MO2-stmn4	MRPHLNO
tp53	MO4-tp53	MRPHLNO

Fish

Fish
AB
AB + MO1-stmn4
AB + MO2-stmn4
rw0130aTg
rw0130aTg + MO1-stmn4
rw0130aTg + MO2-stmn4
rw0130aTg/rw0130aTg
stmn4^{twu0101/twu0101}; rw0130aTg
stmn4^{twu0101/twu0101}; rw0130aTg + MO2-stmn4
stmn4^{twu0102/twu0102}; rw0130aTg

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Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
GFP	EFG	GFP
Kaede	EFG	Kaede

Mapping

Lin et al., 2016 ZDB-PUB-161108-3 PMID:27819330

Stathmin-like 4 is critical for the maintenance of neural progenitor cells in dorsal midbrain of zebrafish larvae

Lin et al., 2016

ZDB-PUB-161108-3
PMID:27819330