Transcriptional regulation of the hypocretin/orexin gene by NR6A1

Biochem Biophys Res Commun. 2010 Dec 10;403(2):178-83. doi: 10.1016/j.bbrc.2010.11.001. Epub 2010 Nov 5.

Abstract

The hypocretin (also known as orexin) neuropeptide system coordinates the regulation of various physiological processes. A reduction in Nr6a1 expression was observed in hypocretin neuron-ablated transgenic mice. To show that prepro-hypocretin transcription is functionally modulated by NR6A1, we performed chromatin immunoprecipitation (ChIP) analysis, double-immunostaining, a luciferase reporter assay, and an in utero electroporation study. ChIP analysis showed that endogenous NR6A1 binds to a putative NR6A1-binding site. Double-immunostaining indicated almost all hypocretin neurons were positive for NR6A1 immunoreactivity. NR6A1 overexpression in SH-SY5Y cells modulated hypocretin promoter activity, an effect that was countered by lacking a putative NR6A1-binding site. Electroporation with Nr6a1 in the foetal hypothalamus promoted hypocretin transcription as compared to GFP-electroporation. These experiments confirmed that NR6A1 works as a regulator for hypocretin transcription.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • DNA Mutational Analysis
  • Gene Expression Regulation*
  • Humans
  • Hypothalamus / metabolism
  • Immunoprecipitation
  • Intracellular Signaling Peptides and Proteins / genetics*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Neuropeptides / genetics*
  • Nuclear Receptor Subfamily 6, Group A, Member 1 / metabolism*
  • Orexins
  • Promoter Regions, Genetic
  • Sequence Deletion
  • Transcription, Genetic

Substances

  • Intracellular Signaling Peptides and Proteins
  • Neuropeptides
  • Nuclear Receptor Subfamily 6, Group A, Member 1
  • Orexins