title: Multiple noncoding exons 1 of nuclear receptors NR4A family (nerve growth factor-induced clone B, Nur-related factor 1 and neuron-derived orphan receptor 1) and NR5A1 (steroidogenic factor 1) in human cardiovascular and adrenal tissues.
abstract: OBJECTIVE
  Nuclear receptors are involved in a wide variety of functions, including aldosteronogenesis. Nuclear receptor families NR4A [nerve growth factor-induced clone B (NGFIB), Nur-related factor 1 (NURR1) and neuron-derived orphan receptor 1 (NOR1)] and NR2F [chicken ovalbumin upstream promoter-transcription factor 1 (COUP-TFI), COUP-TFII and NR2F6) activate, whereas NR5A1 [steroidogenic factor 1 (SF1)] represses CYP11B2 (aldosterone synthase) gene transcription. The present study was undertaken to elucidate the mechanism of differential regulation of nuclear receptors between cardiovascular and adrenal tissues.
  
  
  METHODS
  We collected tissues of artery (n = 9), cardiomyopathy muscle (n = 9), heart muscle (noncardiomyopathy) (n = 6), adrenal gland (n = 9) and aldosterone-producing adenoma (APA) (n = 9). 5'-rapid amplification of cDNA ends (RACE) identified transcription start sites. Multiplex reverse-transcription PCR (RT-PCR) determined use of alternative noncoding exons 1 (ANEs).
  
  
  RESULTS
  In adrenocortical H295R cells, angiotensin II, KCl or cAMP, all stimulated CYP11B2 transcription and NR4A was upregulated, whereas NR2F and NR5A1 were downregulated. 5'-RACE and RT-PCR revealed four ANEs of NGFIB (NR4A1), three of NURR1 (NR4A2), two of NOR1 (NR4A3) and two of SF1 (NR5A1) in cardiovascular and adrenal tissues. Quantitative multiplex RT-PCR showed NR4A and NR5A1 differentially employed multiple ANEs in a tissue-specific manner. The use of ANEs of NGFIB and NURR1 was significantly different between APA and artery. Changes in use of ANEs of NGFIB and NOR1 were observed between cardiomyopathy and noncardiomyopathy. The NR4A mRNA levels in artery were high compared with cardiac and adrenal tissues, whereas the NR5A1 mRNA level in adrenal tissues was extremely high compared with cardiovascular tissues.
  
  
  CONCLUSIONS
  NR4A and NR5A1 genes are complex in terms of alternative promoter use. The use of ANEs may be associated with the pathophysiology of the heart and adrenal gland.
authors.affiliation: Division of Endocrinology and Hypertension, Department of Internal Medicine, Japan. m-demura@med.kanazawa-u.ac.jp
authors.forename: Masashi
authors.initials: M
authors.lastname: Demura
authors.forename: Fen
authors.initials: F
authors.lastname: Wang
authors.forename: Takashi
authors.initials: T
authors.lastname: Yoneda
authors.forename: Shigehiro
authors.initials: S
authors.lastname: Karashima
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Mori
authors.forename: Masashi
authors.initials: M
authors.lastname: Oe
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Kometani
authors.forename: Toshitaka
authors.initials: T
authors.lastname: Sawamura
authors.forename: Yuan
authors.initials: Y
authors.lastname: Cheng
authors.forename: Yuji
authors.initials: Y
authors.lastname: Maeda
authors.forename: Mikio
authors.initials: M
authors.lastname: Namiki
authors.forename: Hidekazu
authors.initials: H
authors.lastname: Ino
authors.forename: Noboru
authors.initials: N
authors.lastname: Fujino
authors.forename: Katsuharu
authors.initials: K
authors.lastname: Uchiyama
authors.forename: Toshinari
authors.initials: T
authors.lastname: Tsubokawa
authors.forename: Masakazu
authors.initials: M
authors.lastname: Yamagishi
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Ono
authors.forename: Hironobu
authors.initials: H
authors.lastname: Sasano
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Demura
authors.forename: Yoshiyu
authors.initials: Y
authors.lastname: Takeda
mesh_terms: D000311:Adrenal Glands / Q000378:metabolism*; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D004247:DNA; D017931:DNA Primers; D005091:Exons; D006801:Humans; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D018160:Receptors, Cytoplasmic and Nuclear; D004247:DNA