PUBLICATION

Three-dimensional structural analysis reveals a Cdk5-mediated kinase cascade regulating hepatic biliary network branching in zebrafish

Authors

Dimri, M., Bilogan, C., Pierce, L.X., Naegele, G., Vasanji, A., Gibson, I., McClendon, A., Tae, K., Sakaguchi, T.F.

ID

ZDB-PUB-170720-3

Date

2017

Source

Development (Cambridge, England) 144: 2595-2605 (Journal)

Registered Authors

Pierce, Lain, Sakaguchi, Takuya

Keywords

Actin dynamics, Biliary atresia, Biliary epithelial cells, Cdk5r1a, Intrahepatic biliary network

MeSH Terms

Zebrafish Proteins/antagonists & inhibitors
Zebrafish Proteins/genetics
Zebrafish Proteins/metabolism*
Cyclin-Dependent Kinase 5/antagonists & inhibitors
Cyclin-Dependent Kinase 5/genetics
Cyclin-Dependent Kinase 5/metabolism*
Morphogenesis/drug effects
Morphogenesis/genetics
Morphogenesis/physiology
Animals, Genetically Modified
Algorithms
Models, Anatomic
Signal Transduction
Gene Knockout Techniques
Animals
Actin Depolymerizing Factors/metabolism
p21-Activated Kinases/metabolism
Larva/growth & development
Larva/metabolism
Bile Ducts, Intrahepatic/enzymology*
Bile Ducts, Intrahepatic/growth & development*
Protein Kinase Inhibitors/pharmacology
Mutation
Lim Kinases/metabolism
Computer Simulation
Zebrafish/genetics
Zebrafish/growth & development*
Zebrafish/metabolism*
Imaging, Three-Dimensional

(all 29)

PubMed

28720653 Full text @ Development

Abstract

The intrahepatic biliary network is a highly branched three-dimensional network lined by biliary epithelial cells, but how its branching patterns are precisely established is not clear. We designed a new computer-based algorithm that quantitatively computes the structural differences of the three-dimensional networks. Utilizing the algorithm, we showed that inhibition of Cyclin-dependent kinase 5 (Cdk5) led to reduced branching in the intrahepatic biliary network in zebrafish. Further, we identified a previously unappreciated downstream kinase cascade regulated by Cdk5. Pharmacological manipulations of this downstream kinase cascade produced a crowded branching defect in the intrahepatic biliary network and influenced actin dynamics in biliary epithelial cells. We generated larvae carrying a mutation in cdk5 regulatory subunit 1a (cdk5r1a), an essential activator of Cdk5. cdk5r1a mutant larvae show similar branching defects as those observed in Cdk5 inhibitor-treated larvae. A small-molecule compound that interferes with the downstream kinase cascade rescued the mutant phenotype. These results provide new insights into branching morphogenesis of the intrahepatic biliary network.

Genes / Markers

Figures

Show all Figures

Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
EGFP	cdk5r1a^lri58/lri58; um14Tg	chemical treatment by environment: LimKi 3	Day 5	bile ductule cholangiocyte	IHC	Fig. 6
EGFP	cdk5r1a^lri58/lri58; um14Tg	control	Day 5	bile ductule cholangiocyte	IHC	Fig. 6
EGFP	um14Tg	control	Day 5	bile ductule cholangiocyte	IHC	Fig. 6
EGFP	um14Tg	chemical treatment by environment: olomoucine	Day 5	bile ductule cholangiocyte	IHC	Fig. 5
EGFP	um14Tg	chemical treatment by environment: cucurbitacin E	Day 5	bile ductule cholangiocyte	IHC	Fig. 4
EGFP	um14Tg	control	Day 5	bile ductule cholangiocyte	IHC	Fig. 4
EGFP	um14Tg	control	Day 5	bile ductule cholangiocyte	IHC	Fig. 5
EGFP	um14Tg	chemical treatment by environment: cucurbitacin E	Day 5	bile ductule cholangiocyte	IHC	Fig. 5
EGFP	um14Tg	chemical treatment by environment: IPA-3	Day 5	bile ductule cholangiocyte	IHC	Fig. 5
EGFP	um14Tg	chemical treatment by environment: LimKi 3	Day 5	bile ductule cholangiocyte	IHC	Fig. 5

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment by environment: IPA-3	Day 5	whole organism Ab2-cfl labeling decreased amount, abnormal	Fig. 5
WT	chemical treatment by environment: LimKi 3	Day 5	whole organism Ab2-cfl labeling decreased amount, abnormal	Fig. 5
WT	chemical treatment by environment: olomoucine	Day 5	whole organism Ab2-cfl labeling increased amount, abnormal	Fig. 5
WT	chemical treatment by environment: olomoucine	Day 5	whole organism Ab5-pak labeling increased amount, abnormal	Fig. 3
cdk5r1a^lri58/lri58; um14Tg	control	Day 5	bile ductule decreased branchiness, abnormal	Fig. 6
cdk5r1a^lri58/lri58; um14Tg	control	Day 5	bile ductule morphogenesis of a branching epithelium decreased occurrence, abnormal	Fig. 6
cdk5r1a^lri58/lri58; um14Tg	control	Day 5	liver and biliary system decreased functionality, abnormal	Fig. 6
cdk5r1a^lri58/lri58; um14Tg	control	Day 5	liver intrahepatic bile duct development decreased process quality, abnormal	Fig. 6
cdk5r1a^lri58/lri58; um14Tg	chemical treatment by environment: LimKi 3	Day 5	bile ductule branchiness, ameliorated	Fig. 6
cdk5r1a^lri58/lri58; um14Tg	chemical treatment by environment: LimKi 3	Day 5	bile ductule morphogenesis of a branching epithelium process quality, ameliorated	Fig. 6

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
lri2Tg	Tg(fabp10a:mCherry-CAAX)	Transgenic Insertion
lri58		Small Deletion	cdk5r1a
um14Tg	Tg(EPV.Tp1-Mmu.Hbb:EGFP)	Transgenic Insertion

Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
cdk5r1a	TALEN1-cdk5r1a	TALEN

Fish

Fish
cdk5r1a^lri58/lri58; um14Tg
um14Tg
WT

Antibodies

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
EGFP	EFG	EGFP
mCherry	EFG	mCherry

Mapping

Dimri et al., 2017 ZDB-PUB-170720-3 PMID:28720653

Three-dimensional structural analysis reveals a Cdk5-mediated kinase cascade regulating hepatic biliary network branching in zebrafish

Dimri et al., 2017

ZDB-PUB-170720-3
PMID:28720653