PUBLICATION

In vivo monitoring of cardiomyocyte proliferation to identify chemical modifiers of heart regeneration

Authors
Choi, W.Y., Gemberling, M., Wang, J., Holdway, J.E., Shen, M.C., Karlstrom, R.O., and Poss, K.D.
ID
ZDB-PUB-130110-29
Date
2013
Source
Development (Cambridge, England)   140(3): 660-666 (Journal)
Registered Authors
Choi, Wen-Yee, Gemberling, Matt, Holdway, Jennifer, Karlstrom, Rolf, Poss, Kenneth D., Wang, Jinhu
Keywords
heart regeneration, chemical screen, cardiomyocyte, cell proliferation, epicardium, endocardium, zebrafish
MeSH Terms
  • Catechols/pharmacology
  • Female
  • Animals
  • Heart/embryology
  • Heart/physiology*
  • Isoquinolines/pharmacology
  • Myocytes, Cardiac/cytology*
  • Myocytes, Cardiac/drug effects
  • Myocytes, Cardiac/metabolism
  • Male
  • Biomarkers/metabolism
  • Transgenes
  • Cell Proliferation*
  • Animals, Genetically Modified/embryology
  • Animals, Genetically Modified/metabolism
  • Animals, Genetically Modified/physiology
  • Embryo, Nonmammalian/cytology
  • Embryo, Nonmammalian/drug effects
  • Embryo, Nonmammalian/physiology
  • Cell Count
  • Thiophenes/pharmacology
  • Hedgehog Proteins/agonists
  • Hedgehog Proteins/genetics
  • Hedgehog Proteins/metabolism
  • Regeneration*
  • Ubiquitination
  • Cyclohexylamines/pharmacology
  • Recombinant Fusion Proteins/genetics
  • Recombinant Fusion Proteins/metabolism
  • Zebrafish/genetics
  • Zebrafish/injuries
  • Zebrafish/physiology
  • Signal Transduction
  • High-Throughput Screening Assays/methods*
  • Transforming Growth Factor beta/genetics
  • Transforming Growth Factor beta/metabolism
(all 36)
PubMed
23293297 Full text @ Development
Abstract

Adult mammalian cardiomyocytes have little capacity to proliferate in response to injury, a deficiency that underlies the poor regenerative ability of human hearts after myocardial infarction. By contrast, zebrafish regenerate heart muscle after trauma by inducing proliferation of spared cardiomyocytes, providing a model for identifying manipulations that block or enhance these events. Although direct genetic or chemical screens of heart regeneration in adult zebrafish present several challenges, zebrafish embryos are ideal for high-throughput screening. Here, to visualize cardiomyocyte proliferation events in live zebrafish embryos, we generated transgenic zebrafish lines that employ fluorescent ubiquitylation-based cell cycle indicator (FUCCI) technology. We then performed a chemical screen and identified several small molecules that increase or reduce cardiomyocyte proliferation during heart development. These compounds act via Hedgehog, Insulin-like growth factor or Transforming growth factor β signaling pathways. Direct examination of heart regeneration after mechanical or genetic ablation injuries indicated that these pathways are activated in regenerating cardiomyocytes and that they can be pharmacologically manipulated to inhibit or enhance cardiomyocyte proliferation during adult heart regeneration. Our findings describe a new screening system that identifies molecules and pathways with the potential to modify heart regeneration.

Genes / Markers
Figures
Figure Gallery (8 images)
Show all Figures
Expression
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay Figure
EGFPsb15Tgstandard conditionsAdultNot DetectedIHC
EGFPsb15Tgphysical alteration: anatomical structureAdultIHC
EGFPumz23Tgstandard conditionsAdultNot DetectedIHC
EGFPumz23Tgphysical alteration: anatomical structureAdultIHC
igf2bWTstandard conditionsAdultNot DetectedISH
igf2bWTphysical alteration: anatomical structureAdultISH
mCherrypd57Tg/+; pd58Tg/+chemical treatment: pharmaceuticalDay 4IFL
mCherrypd57Tg/+; pd58Tg/+chemical treatment: pharmaceuticalDay 4IFL
mCherrypd57Tg/+; pd58Tg/+controlDay 4IFL
mCherrypd57Tg/+; pd58Tg/+standard conditionsDay 4IFL
1 - 10 of 16
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Phenotype
Fish Conditions Stage Phenotype Figure
pd57Tg/+; pd58Tg/+chemical treatment: CyclopamineDay 4
pd57Tg/+; pd58Tg/+chemical treatment: pharmaceuticalDay 4
pd57Tg/+; pd58Tg/+chemical treatment: inhibitorDay 4
pd57Tg/+; pd58Tg/+chemical treatment: pharmaceuticalDay 4
1 - 4 of 4
Show
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
f2TgTransgenic Insertion
    pd10TgTransgenic Insertion
      pd36TgTransgenic Insertion
        pd57TgTransgenic Insertion
          pd58TgTransgenic Insertion
            sb15TgTransgenic Insertion
              umz23TgTransgenic Insertion
                1 - 7 of 7
                Show
                Human Disease / Model
                No data available
                Sequence Targeting Reagents
                No data available
                Fish
                Fish
                f2Tg
                pd10Tg; pd36Tg
                pd57Tg/+; pd58Tg/+
                sb15Tg
                umz23Tg
                WT
                1 - 6 of 6
                Show
                Antibodies
                Orthology
                No data available
                Engineered Foreign Genes
                Marker Marker Type Name
                CreEFGCre
                DipToxEFGDipTox
                DsRedEFGDsRed
                DsRed2EFGDsRed2
                EGFPEFGEGFP
                GFPEFGGFP
                mCherryEFGmCherry
                VenusEFGVenus
                1 - 8 of 8
                Show
                Mapping
                No data available
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                Citation
                Choi, W.Y., Gemberling, M., Wang, J., Holdway, J.E., Shen, M.C., Karlstrom, R.O., and Poss, K.D. (2013) In vivo monitoring of cardiomyocyte proliferation to identify chemical modifiers of heart regeneration. Development (Cambridge, England). 140(3):660-666.