Transcriptional regulation of the hypocretin/orexin gene by NR6A1.
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.
Department of Sleep Disorders Research, Tokyo Institute of Psychiatry, Tokyo, Japan. tanaka-ss@igakuken.or.jp
Susumu
S
Tanaka
Tohru
T
Kodama
Takashi
T
Nonaka
Hiromi
H
Toyoda
Makoto
M
Arai
Miyuki
M
Fukazawa
Yoshiko
Y
Honda
Makoto
M
Honda
Emmanuel
E
Mignot
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004252:DNA Mutational Analysis; D005786:Gene Expression Regulation; D006801:Humans; D007031:Hypothalamus / Q000378:metabolism; D047468:Immunoprecipitation; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000235:genetics*; D057136:Nuclear Receptor Subfamily 6, Group A, Member 1 / Q000378:metabolism*; D000068797:Orexins; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D014158:Transcription, Genetic
D047908:Intracellular Signaling Peptides and Proteins; D009479:Neuropeptides; D057136:Nuclear Receptor Subfamily 6, Group A, Member 1; D000068797:Orexins