PUBLICATION

Loss of Dnmt1 catalytic activity reveals multiple roles for DNA methylation during pancreas development and regeneration

Authors
Anderson, R.M., Bosch, J.A., Goll, M.G., Hesselson, D., Dong, P.D., Shin, D., Chi, N.C., Shin, C.H., Schlegel, A., Halpern, M., and Stainier, D.Y.
ID
ZDB-PUB-090731-2
Date
2009
Source
Developmental Biology   334(1): 213-223 (Journal)
Registered Authors
Anderson, Ryan, Chi, Neil C., Dong, P. Duc, Goll, Mary, Halpern, Marnie E., Hesselson, Daniel, Schlegel, Amnon, Shin, Chong, Shin, Donghun, Stainier, Didier
Keywords
Methylation, Dnmt1, Pancreas, Epigenetics, Regeneration, Zebrafish, ENU mutagenesis, Mutant, Beta cells, Liver
MeSH Terms
  • Cell Survival
  • DNA Methylation
  • Endocrine Cells/metabolism
  • Molecular Sequence Data
  • Zebrafish/metabolism
  • DNA (Cytosine-5-)-Methyltransferases/genetics*
  • DNA (Cytosine-5-)-Methyltransferases/metabolism
  • Insulin-Secreting Cells/cytology
  • Insulin-Secreting Cells/metabolism
  • Animals
  • Base Sequence
  • Zebrafish Proteins/genetics*
  • Zebrafish Proteins/metabolism
  • Fluorescent Antibody Technique
  • Pancreas/cytology*
  • Pancreas/growth & development
  • Regeneration/physiology*
  • Amino Acid Sequence
(all 18)
PubMed
19631206 Full text @ Dev. Biol.
Abstract
Developmental mechanisms regulating gene expression and the stable acquisition of cell fate direct cytodifferentiation during organogenesis. Moreover, it is likely that such mechanisms could be exploited to repair or regenerate damaged organs. DNA methyltransferases (Dnmts) are enzymes critical for epigenetic regulation, and are used in concert with histone methylation and acetylation to regulate gene expression and maintain genomic integrity and chromosome structure. We carried out two forward genetic screens for regulators of endodermal organ development. In the first, we screened for altered morphology of developing digestive organs, while in the second we screed for the lack of terminally differentiated cell types in the pancreas and liver. From these screens, we identified two mutant alleles of zebrafish dnmt1. Both lesions are predicted to eliminate dnmt1 function; one is a missense mutation in the catalytic domain and the other is a nonsense mutation that eliminates the catalytic domain. In zebrafish dnmt1 mutants, the pancreas and liver form normally, but begin to degenerate after 84 hours post fertilization (hpf). Acinar cells are nearly abolished through apoptosis by 100 hpf, though neither DNA replication, nor entry into mitosis are halted in the absence of detectable Dnmt1. However, endocrine cells and ducts are largely spared. Surprisingly, dnmt1 mutants and dnmt1 morpholino-injected larvae show increased capacity for pancreatic beta cell regeneration in an inducible model of pancreatic beta cell ablation. Thus, our data suggest that Dnmt1 is dispensable for pancreatic duct or endocrine cell formation, but not for acinar cell survival. In addition, Dnmt1 may influence the differentiation of pancreatic beta cell progenitors or the reprogramming of cells toward the pancreatic beta cell fate.
Genes / Markers
Figures
Figure Gallery (6 images)
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Expression
Phenotype
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
gz4TgTransgenic Insertion
    gz15TgTransgenic Insertion
      ia3TgTransgenic Insertion
        jh1TgTransgenic Insertion
          m1018TgTransgenic Insertion
            s843TgTransgenic Insertion
              s854TgTransgenic Insertion
                s872
                  MNV
                  s892TgTransgenic Insertion
                    s904
                      Point Mutation
                      1 - 10 of 10
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                      Human Disease / Model
                      No data available
                      Sequence Targeting Reagents
                      Target Reagent Reagent Type
                      dnmt1MO4-dnmt1MRPHLNO
                      tp53MO4-tp53MRPHLNO
                      1 - 2 of 2
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                      Fish
                      Antibodies
                      Orthology
                      No data available
                      Engineered Foreign Genes
                      Marker Marker Type Name
                      CFPEFGCFP
                      DsRedEFGDsRed
                      EGFPEFGEGFP
                      GFPEFGGFP
                      NTREFGNTR
                      RFPEFGRFP
                      1 - 6 of 6
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                      Mapping
                      Entity Type Entity Symbol Location
                      Features872Chr: 3 Details
                      Features904Chr: 3 Details
                      GENEangptl6Chr: 3 Details
                      GENEdnmt1Chr: 3 Details
                      GENEeif3gChr: 3 Details
                      GENEppanChr: 3 Details
                      GENEs1pr2Chr: 3 Details
                      1 - 7 of 7
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