Comprehensive identification of the Pet-1-target genes by chromatin immunoprecipitation assays in RN46A cells derived from the rat raphe nucleus.

• Project/Area Number: 24659548
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Psychiatric science
• Research Institution: St. Marianna University School of Medicine
• Principal Investigator: MATSUI Hiroaki 聖マリアンナ医科大学, 医学(系)研究科(研究院), 教授 (90181685)
• Research Collaborator: NAWA Yukino 聖マリアンナ医科大学, 医学(系)研究科(研究院), 助教 (10549786)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2012: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: セロトニン神経系 / クロマチン免疫沈降法 / ラット縫線核由来RN46A細胞 / セロトニン神経細胞特異的転写因子Pet-1 / Pet-1標的遺伝子 / 神経特異的抑制因子 / 転写因子 / 遺伝子発現 / クロマチン沈降法

Research Abstract

To identify Pet-1 (serotonergic neuron-specific transcription factor)-binding sites in rat raphe nucleus-derived RN46A cells, chromatin-immunoprecipitation (ChIP) assays were performed. It was revealed that Pet-1 binding was inhibited through the neuron-restrictive silencer factor(NRSF)-NRSF binding element(NRSE) system or the potential repressor proteins independent of NRSF. Additionally, the detection of Pet-1 binding to target genes was occasionally difficult even under conditions where Pet-1 was overexpressed. Taken together, to exclude the function of repressor proteins including NRSF may be necessary to identify the Pet-1 binding to its target genes by ChIP assays in RN46A cells. To overcome these difficulties should be required to use RN46A cells as a proper model of central serotonergic neurons and characterize the Pet-1-mediated transcriptional network in serotonergic neurons.

Research Products

• [Journal Article] Tryptophan hydroxylase 2 (TPH2) in a neuronal cell line : modulation by cell differentiation and NRSF/ rest activity (2012)
  • Author(s): Gentile MT, Nawa Y, Lunardi G, Florio T, Matsui H, Colucci-D'Amato L
  • Journal Title: Journal of Neurochemistry Volume: Vol. 123、No.6 Issue: 6 Pages: 963-970
  • DOI: 10.1111/jnc.12004
  • Peer Reviewed
• [Presentation] ヒトセロトニン4受容体遺伝子発現調節機構 : ERG、EGRおよびNF1による調節 (2013)
  • Author(s): 那和雪乃
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 兵庫県神戸市
  • Year and Date: 2013-12-04
• [Presentation] Transcriptional regulation of the human serotonin type 4(5-ht4) receptor gene : identification and characterization of novel promoter and enhancer (2013)
  • Author(s): Nawa Y
  • Organizer: Annual Meeting Neuroscience 2013
  • Place of Presentation: San Diego、U.S.A
  • Year and Date: 2013-11-13
• [Presentation] Transcriptional regulation of the human serotonin type 4(5-ht4) receptor gene:identification and characterization of novel promoter and enhancer (2013)
  • Author(s): Yukino Nawa, Masanori Ootaki, Rie Kuwabara, Tomoko Hiroi, Ryoya Takahashi, Hiroaki Matsui
  • Organizer: 北米神経科学会
  • Place of Presentation: サンディエゴ（米国）
• [Presentation] ヒトセロトニン4受容体遺伝子発現調節機構：ERG、EGRおよびNF1による調節 (2013)
  • Author(s): 那和雪乃、大滝正則、桑原理恵、廣井朋子、高橋良哉、松井宏晃
  • Organizer: 第36回日本分子生物学会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
• [Presentation] Transcriptional regulation of the human tryptophan hydroxylase-2 gene by Lmx1b in RN46A cells (2012)
  • Author(s): Nawa Y
  • Organizer: Annual Meeting Neuroscience 2012
  • Place of Presentation: New Orleans、U.S.A
  • Year and Date: 2012-10-14
• [Presentation] Transcriptional regulation of the human tryptophan hydroxylase-2 gene by Lmx1b in RN46A cells (2012)
  • Author(s): Yukino Nawa
  • Organizer: 北米神経科学会
  • Place of Presentation: ニューオリンズ（米国）
• [Remarks] 聖マリアンナ医科大学大学院脳情報制御医学分野
  • URL: http://www.marianna-u.ac.jp/isotope/facilities/12541/012540.html