PUBLICATION

Cell-autonomous regulation of epithelial cell quiescence by calcium channel Trpv6

Authors

Xin, Y., Malick, A., Hu, M., Liu, C., Batah, H., Xu, H., Duan, C.

ID

ZDB-PUB-190919-2

Date

2019

Source

eLIFE 8: (Journal)

Registered Authors

Duan, Cunming

Keywords

developmental biology, zebrafish

MeSH Terms

Epithelial Cells/physiology*
Calcium Channels/metabolism*
Zebrafish Proteins/deficiency
Zebrafish Proteins/metabolism
Signal Transduction
Gene Deletion
Humans
Animals
Calcium/metabolism
TRPV Cation Channels/deficiency
TRPV Cation Channels/metabolism*
Intercellular Signaling Peptides and Proteins/metabolism
Cell Proliferation*
Zebrafish

(all 14)

PubMed

31526479 Full text @ Elife

Abstract

Epithelial homeostasis and regeneration require a pool of quiescent cells. How the quiescent cells are established and maintained is poorly understood. Here we report that Trpv6, a cation channel responsible for epithelial Ca²⁺ absorption, functions as a key regulator of cellular quiescence. Genetic deletion and pharmacological blockade of Trpv6 promoted zebrafish epithelial cells to exit from quiescence and re-enter the cell cycle. Reintroducing Trpv6, but not its channel dead mutant, restored the quiescent state. Ca²⁺ imaging showed that Trpv6 is constitutively open in vivo. Mechanistically, Trpv6-mediated Ca²⁺ influx maintained the quiescent state by suppressing insulin-like growth factor (IGF)-mediated Akt-Tor and Erk signaling. In zebrafish epithelia and human colon carcinoma cells, Trpv6/TRPV6 elevated intracellular Ca²⁺ levels and activated PP2A, which down-regulated IGF signaling and promoted the quiescent state. Our findings suggest that Trpv6 mediates constitutive Ca²⁺ influx into epithelial cells to continuously suppress growth factor signaling and maintain the quiescent state.

Genes / Markers

Figures

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Expression

Gene	Fish	Conditions	Stage	Anatomy	Assay	Figure
GCaMP	mi603Tg/+	chemical treatment: ionomycin	Protruding-mouth	NaK ionocyte	IFL	Figure 4
GCaMP	mi603Tg/+	control	Protruding-mouth	NaK ionocyte	IFL	Figure 4
GCaMP	trpv6^mi606/mi606; mi603Tg/+	chemical treatment: ionomycin	Protruding-mouth	NaK ionocyte	IFL	Figure 4
GCaMP	trpv6^mi606/mi606; mi603Tg/+	control	Protruding-mouth	NaK ionocyte	IFL	Figure 4
GFP	mi602Tg	control	Protruding-mouth	NaK ionocyte	IFL	Figure 2
GFP	mi602Tg	control	Day 4	NaK ionocyte	IFL	Figure 2
GFP	mi602Tg	control	Day 5	NaK ionocyte	IFL	Figure 2
GFP	trpv6^mi604/mi604; mi602Tg	control	Day 5	NaK ionocyte	IFL	Figure 2
GFP	trpv6^mi604/mi604; mi602Tg	control	Protruding-mouth	NaK ionocyte	IFL	Figure 2
GFP	trpv6^mi604/mi604; mi602Tg	control	Day 4	NaK ionocyte	IFL	Figure 2

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Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	chemical treatment: 2-[N-[2-(acetyloxymethoxy)-2-oxoethyl]-2-[2-[2-[bis[2-(acetyloxymethoxy)-2-oxoethyl]amino]phenoxy]ethoxy]anilino]acetic acid acetyloxymethyl ester	Day 4	integument ab5-akt labeling increased distribution, abnormal	Figure 4
WT	chemical treatment: 2-[N-[2-(acetyloxymethoxy)-2-oxoethyl]-2-[2-[2-[bis[2-(acetyloxymethoxy)-2-oxoethyl]amino]phenoxy]ethoxy]anilino]acetic acid acetyloxymethyl ester	Day 5	NaK ionocyte increased amount, abnormal	Figure 4
mi602Tg	chemical treatment: calyculin A	Day 4	whole organism ab5-akt labeling increased amount, abnormal	Fig. 5
mi602Tg	chemical treatment: calyculin A	Day 4	whole organism ab5-akt labeling increased distribution, abnormal	Fig. 5
mi602Tg	chemical treatment: calyculin A	Day 5	NaK ionocyte increased amount, abnormal	Fig. 5
mi602Tg	chemical treatment: calyculin A	Day 5	NaK ionocyte increased amount, abnormal	Fig. 5 supplement 1
mi602Tg	chemical treatment: BMS-754807, chemical treatment: calyculin A	Day 4	whole organism ab5-akt labeling amount, ameliorated	Fig. 5
mi602Tg	chemical treatment: BMS-754807, chemical treatment: calyculin A	Day 5	NaK ionocyte amount, ameliorated	Fig. 5
mi602Tg	chemical treatment: gadolinium trichloride	Day 5	NaK ionocyte increased amount, abnormal	Figure 2—figure supplement 2.
mi602Tg	chemical treatment: gadolinium trichloride	Day 5	NaK ionocyte increased amount, abnormal	Figure 2

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Mutations / Transgenics

Allele	Construct	Type	Affected Genomic Region
mi602Tg	TgBAC(igfbp5a:GFP)	Transgenic Insertion
mi603Tg	TgBAC(igfbp5a:GCaMP7)	Transgenic Insertion
mi604		Small Deletion	trpv6
mi605		Indel	trpv6
mi606		Indel	trpv6

1 - 5 of 5

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Human Disease / Model

Sequence Targeting Reagents

Target	Reagent	Reagent Type
ppp2caa	CRISPR1-ppp2caa	CRISPR
ppp2cab	CRISPR1-ppp2cab	CRISPR
ppp2cb	CRISPR1-ppp2cb	CRISPR
trpv6	CRISPR1-trpv6	CRISPR

Fish

Fish
mi602Tg
mi602Tg + CRISPR1-ppp2caa + CRISPR1-ppp2cab + CRISPR1-ppp2cb
mi603Tg/+
trpv6^mi604/mi604; mi602Tg
trpv6^mi604/+; mi602Tg
trpv6^mi605/mi605; mi602Tg
trpv6^mi605/+; mi602Tg
trpv6^mi606/mi606; mi603Tg/+
WT

1 - 9 of 9

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Antibodies

Name	Type	Isotypes	Host Organism
Ab2-rps6	monoclonal	IgG	Rabbit
Ab5-akt	monoclonal		Rabbit
Ab9-mapk	monoclonal	IgG	Rabbit

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
GCaMP	EFG	GCaMP
GFP	EFG	GFP

Mapping

Xin et al., 2019 ZDB-PUB-190919-2 PMID:31526479

Cell-autonomous regulation of epithelial cell quiescence by calcium channel Trpv6

Xin et al., 2019

ZDB-PUB-190919-2
PMID:31526479