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hnf1b genes in zebrafish hindbrain development.

Seong-Kyu Choe1, Nicolas Hirsch, Xiaolan Zhang

  • 1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, Massachusetts 01605-2324, USA.

Zebrafish
|October 24, 2008
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Summary

Hepatocyte nuclear factor 1-beta (Hnf1b) is essential for hindbrain development. Studies show Hnf1b is required for gene expression in rhombomeres 5 and 6, but this can be induced elsewhere in the embryo.

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Area of Science:

  • Developmental biology
  • Genetics
  • Molecular biology

Background:

  • The transcription factor Hepatocyte nuclear factor 1-beta (Hnf1b) plays a role in hindbrain formation.
  • Its precise requirement in rhombomeres (r) 5 and 6 during hindbrain development is not fully understood.

Purpose of the Study:

  • To investigate the essential role of Hnf1b in the development of rhombomeres 5 and 6.
  • To analyze the function of specific Hnf1b alleles and their impact on gene expression.

Main Methods:

  • Examination of Hnf1b retroviral insertion alleles (hnf1b(hi2169) and hnf1b(hi1843)).
  • Identification and knockdown of a novel virus-Hnf1b fusion transcript.
  • Microinjection of hoxb1b mRNA into mutant embryos.

Main Results:

  • The hnf1b(hi2169) allele exhibits variable residual gene expression due to a virus-Hnf1b fusion transcript, not full-length Hnf1b or Hnf1bl.
  • Morpholino knockdown of the fusion transcript led to near-undetectable r5 gene expression.
  • The hnf1b(hi1843) allele showed a more severe phenotype with minimal r5/r6 gene expression.
  • Hoxb1b mRNA could induce ectopic gene expression anterior to the hindbrain but not restore r5/r6 gene expression in mutants.

Conclusions:

  • Hnf1b is indispensable for proper gene expression in hindbrain rhombomeres 5 and 6.
  • The hnf1b(hi2169) allele is hypomorphic compared to hnf1b(hi1843).
  • While Hnf1b is required in the hindbrain, gene expression can be induced independently of Hnf1b in anterior regions.