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Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization
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Dnmt3 and G9a cooperate for tissue-specific development in zebrafish.

Kunal Rai1, Itrat F Jafri1, Stephanie Chidester2

  • 1From the Departments of Oncological Sciences, University of Utah, Salt Lake City, Utah 84112; Departments of Huntsman Cancer Institute, University of Utah, Salt Lake City, Utah 84112 and; Departments of Howard Hughes Medical Institute, University of Utah, Salt Lake City, Utah 84112.

The Journal of Biological Chemistry
|December 1, 2009
PubMed
Summary

DNA methyltransferase 3 (Dnmt3) is crucial for zebrafish neurogenesis and brain development. Specific DNMT-histone methyltransferase networks regulate cell fate, highlighting tissue-specific roles in development.

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Area of Science:

  • Developmental Biology
  • Epigenetics
  • Molecular Biology

Background:

  • DNA methylation is essential for embryonic and tissue-specific development.
  • The distinct roles of DNA methyltransferases (DNMTs) in tissue-specific development and their targets are not fully understood.

Purpose of the Study:

  • To investigate the roles of Dnmt3 and Dnmt1 in zebrafish development.
  • To identify Dnmt3-specific target genes and understand its regulatory mechanisms.

Main Methods:

  • Antisense morpholino knockdown of Dnmt3 and Dnmt1 in zebrafish.
  • Phenotypic analysis of knockdown effects on various organs, particularly the brain and retina.
  • Identification and validation of Dnmt3 target genes, including lef1.
  • Investigating the interplay between Dnmt3, G9a, and Suv39h1 in regulating gene expression and development.

Main Results:

  • Dnmt3 is essential for neurogenesis, with its absence causing severe brain and retina defects, while other organs remain unaffected.
  • Dnmt1 and Dnmt3 exhibit distinct functions and cannot compensate for each other's deficiency.
  • lef1 was identified as a Dnmt3-specific target gene, crucial for neurogenesis.
  • Dnmt3 cooperates with G9a, but not Suv39h1, in regulating lef1 and tissue-specific development, forming specific DNMT-histone methyltransferase networks.

Conclusions:

  • Dnmt3 plays a critical, tissue-specific role in neurogenesis.
  • Specific DNMT-histone methyltransferase networks, such as Dnmt3-G9a, are utilized to silence cell fate regulators in a tissue-specific manner.
  • This study elucidates the distinct functions of DNMTs and their collaboration with histone methyltransferases in developmental processes.