V-ATPase proton pumping activity is required for adult zebrafish appendage regeneration

Joana Monteiro; Rita Aires; Jörg D Becker; António Jacinto; Ana C Certal; Joaquín Rodríguez-León

doi:10.1371/journal.pone.0092594

V-ATPase proton pumping activity is required for adult zebrafish appendage regeneration

PLoS One. 2014 Mar 26;9(3):e92594. doi: 10.1371/journal.pone.0092594. eCollection 2014.

Authors

Joana Monteiro¹, Rita Aires², Jörg D Becker², António Jacinto³, Ana C Certal⁴, Joaquín Rodríguez-León⁵

Affiliations

¹ Instituto Gulbenkian de Ciência, Oeiras, Portugal; Instituto de Medicina Molecular, Faculdade de Medicina de Lisboa, Lisboa, Portugal.
² Instituto Gulbenkian de Ciência, Oeiras, Portugal.
³ Centro de Estudos de Doenças Crónicas, Faculdade de Ciências Médicas, Lisboa, Portugal.
⁴ Instituto Gulbenkian de Ciência, Oeiras, Portugal; Champalimaud Foundation, Lisboa, Portugal.
⁵ Instituto Gulbenkian de Ciência, Oeiras, Portugal; Department de Anatomía Humana, Biología Celular y Zoología, Facultad de Medicina, Universidad de Extremadura, Badajoz, Spain.

Abstract

The activity of ion channels and transporters generates ion-specific fluxes that encode electrical and/or chemical signals with biological significance. Even though it is long known that some of those signals are crucial for regeneration, only in recent years the corresponding molecular sources started to be identified using mainly invertebrate or larval vertebrate models. We used adult zebrafish caudal fin as a model to investigate which and how ion transporters affect regeneration in an adult vertebrate model. Through the combined use of biophysical and molecular approaches, we show that V-ATPase activity contributes to a regeneration-specific H+ ef`flux. The onset and intensity of both V-ATPase expression and H+ efflux correlate with the different regeneration rate along the proximal-distal axis. Moreover, we show that V-ATPase inhibition impairs regeneration in adult vertebrate. Notably, the activity of this H+ pump is necessary for aldh1a2 and mkp3 expression, blastema cell proliferation and fin innervation. To the best of our knowledge, this is the first report on the role of V-ATPase during adult vertebrate regeneration.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amputation, Surgical
Animal Fins / innervation
Animal Fins / physiology
Animals
Cell Proliferation / drug effects
Gene Knockdown Techniques
Larva / physiology
Morpholinos / pharmacology
Proton Pumps / metabolism*
Protons
Regeneration / drug effects
Regeneration / physiology*
Up-Regulation / drug effects
Vacuolar Proton-Translocating ATPases / antagonists & inhibitors
Vacuolar Proton-Translocating ATPases / metabolism*
Zebrafish / physiology*

Substances

Morpholinos
Proton Pumps
Protons
Vacuolar Proton-Translocating ATPases

Grants and funding

JJM was supported by a fellowship (SFRH/BD/45131/2008) from Fundação para a Ciência e Tecnologia (FCT), Portugal. ACC was supported by a postdocoral fellowship (SFRH/BPD/29957/2006) from Fundação para a Ciência e Tecnologia (FCT), Portugal. This work was supported by the following grants: european FP6-Cells into Organs, FCT (Grant PTDC/BIA-BCM/100867/2008 and POCTI-ISFL-4-664) and Ministerio de Ciencia e Innovación, Spain (Subprograma Ramon y Cajal, reference RYC-2008-02753 and grant BFU2009-12279). JRL is supported by the Subprogram Ramón y Cajal from Ministerio de Ciencia e Innovación, Spain (RYC-2008-02753). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.