PUBLICATION

Two T-box genes play independent and cooperative roles to regulate morphogenesis of ciliated Kupffer's vesicle in zebrafish

Authors

Amack, J.D., Wang, X., and Yost, H.J.

ID

ZDB-PUB-070907-29

Date

2007

Source

Developmental Biology 310(2): 196-210 (Journal)

Registered Authors

Amack, Jeffrey, Yost, H. Joseph

Keywords

Left–right asymmetry; Dorsal forerunner cells; Kupffer's vesicle; T-box genes; Tbx16/spadetail; No tail/brachyury; Cilia; Nodal flow

MeSH Terms

Mesoderm/cytology*
Mesoderm/physiology
Zebrafish/embryology
Zebrafish/physiology*
Body Patterning/physiology
Fetal Proteins
Protein Kinase C/metabolism
Cell Differentiation/physiology
Cilia/physiology
T-Box Domain Proteins/genetics
T-Box Domain Proteins/physiology*
Embryo, Nonmammalian/physiology
Animals
Epithelium/physiology
Zebrafish Proteins/genetics
Zebrafish Proteins/physiology*

(all 16)

PubMed

17765888 Full text @ Dev. Biol.

Abstract

The brain, heart and gastro-intestinal tract develop distinct left-right (LR) asymmetries. Asymmetric cilia-dependent fluid flow in the embryonic node in mouse, Kupffer's vesicle in zebrafish, notochordal plate in rabbit and gastrocoel roof plate in frog appears to be a conserved mechanism that directs LR asymmetric gene expression and establishes the orientation of organ asymmetry. However, the cellular processes and genetic pathways that control the formation of these essential ciliated structures are unknown. In zebrafish, migratory dorsal forerunner cells (DFCs) give rise to Kupffer's vesicle (KV), a ciliated epithelial sheet that forms a lumen and generates fluid flow. Using the epithelial marker atypical Protein Kinase C (aPKC) and other markers to analyze DFCs and KV cells, we describe a multi-step process by which DFCs form a functional KV. Using mutants and morpholinos, we show that two T-box transcription factors-No tail (Ntl)/Brachyury and Tbx16/Spadetail-cooperatively regulate an early step of DFC mesenchyme to epithelial transition (MET) and KV cell specification. Subsequently, each transcription factor independently controls a distinct step in KV formation: Tbx16 regulates apical clustering of KV cells and Ntl is necessary for KV lumen formation. By targeting morpholinos to DFCs, we show that these cell autonomous functions in KV morphogenesis are necessary for LR patterning throughout the embryo.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Qualifier	Anatomy	Assay	Figure
dand5	AB	standard conditions	10-13 somites		tail bud	ISH	Fig. 3
dand5	tbx16^b104/b104 (AB)	standard conditions	10-13 somites		tail bud	ISH	Fig. 3
dand5	tbxta^b160/b160 (AB)	standard conditions	10-13 somites	Not Detected	tail bud	ISH	Fig. 3
lft1	AB	standard conditions	20-25 somites		diencephalon	ISH	Fig. 5
lft1	AB + MO3-tbx16	standard conditions	20-25 somites		diencephalon	ISH	Fig. 5
lft2	AB	standard conditions	20-25 somites		heart rudiment	ISH	Fig. 5
lft2	AB + MO3-tbx16	standard conditions	20-25 somites		heart rudiment	ISH	Fig. 5
myod1	AB	standard conditions	Prim-5		somite	ISH	Fig. S3
myod1	AB + MO3-tbx16	standard conditions	Prim-5		somite	ISH	Fig. S3
sb:cb81	AB	standard conditions	75%-epiboly		forerunner cell group	ISH	text only

1 - 10 of 67

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + MO1-tbxta	standard conditions	1-4 somites to 5-9 somites	Kupffer's vesicle unlumenized, abnormal	Fig. 7
AB + MO1-tbxta	standard conditions	Prim-5	post-vent region shape, abnormal	Fig. 8
AB + MO1-tbxta	standard conditions	Prim-5	trunk morphology, abnormal	Fig. 8
AB + MO1-tbxta + MO3-tbx16	standard conditions	5-9 somites	Kupffer's vesicle ciliated cell absent, abnormal	Fig. 8
AB + MO1-tbxta + MO3-tbx16	standard conditions	Prim-5	post-vent region shape, abnormal	Fig. 8
AB + MO1-tbxta + MO3-tbx16	standard conditions	Prim-5	trunk morphology, abnormal	Fig. 8
AB + MO3-tbx16	standard conditions	5-9 somites	Kupffer's vesicle ciliated cell present, normal	Fig. 5
AB + MO3-tbx16	standard conditions	5-9 somites	Kupffer's vesicle decreased size, abnormal	Fig. 5
AB + MO3-tbx16	standard conditions	5-9 somites	Kupffer's vesicle malformed, abnormal	Fig. 5
AB + MO3-tbx16	standard conditions	20-25 somites	determination of left/right symmetry disrupted, abnormal	Fig. 5

1 - 10 of 37

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
b104		Indel	tbx16
b160		Indel	tbxta

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
tbx16	MO3-tbx16	MRPHLNO
tbxta	MO1-tbxta	MRPHLNO

Fish

Fish
AB
AB + MO1-tbxta
AB + MO1-tbxta + MO3-tbx16
AB + MO3-tbx16
tbx16^b104/b104 (AB)
tbx16^b104/b104; tbxta^b160/b160 (AB)
tbxta^b160/b160 (AB)

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Amack et al., 2007 ZDB-PUB-070907-29 PMID:17765888

Two T-box genes play independent and cooperative roles to regulate morphogenesis of ciliated Kupffer's vesicle in zebrafish

Amack et al., 2007

ZDB-PUB-070907-29
PMID:17765888