Molecular mechanism of neural crest cell differentiation, proliferation and death

• Project/Area Number: 16590240
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pathological medical chemistry
• Research Institution: Chiba University
• Principal Investigator: HATANO Masahiko Chiba University, Biomedical Research Center, Associate Professor, バイオメディカル研究センター, 助教授 (20208523)
• Co-Investigator(Kenkyū-buntansha): FUJIMURA Lisa Chiba University, Biomedical Research Center, Assistant Prof., バイオメディカル研究センター, 助手 (30376363) ; TOKUHISA Takeshi Chiba University, Graduate School of Medicine, Professor, 大学院・医学研究院, 教授 (20134364) ; ARIMA Masafumi Chiba University, Graduate School of Medicine, Lecturer, 大学院・医学研究院, 講師 (00202763) ; SAKAMOTO Akemi Chiba University, Graduate School of Medicine, Assistant Prof., 大学院・医学研究院, 助手 (90359597)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2005: ¥1,700,000 (Direct Cost: ¥1,700,000); Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
• Keywords: Ncx / Neural crest cell / Nczf / Target genes / Neural cell death / Transcriptional repressor / DNA binding / Hirschsprung related diseases / Zinc finger protein / DNA結合配列 / 腸管神経節 / 遺伝子欠損マウス / 転写因子 / クロマチン免疫沈降 / ノックアウトマウス

Research Abstract

The murine Ncx (Tlx2,Enx,Hox11L1) gene is expressed in neural crest derived tissues and regulates neuronal cell death in enteric neurons. We have used chromatin immunoprecipitation method to screen for target genes for Ncx. This screen led to the identification of novel gene termed Nczf. Nczf contains an N-terminal KRAB box domain and 11 Kruppel C2H2 type zinc finger motifs at C terminus. Promoter region of the Nczf gene contains 5 consensus sequences for Ncx binding motifs. Transient transfection assays of the 5'-flanking region fused to the luciferase reporter gene with various amount of Ncx expression vector showed dose dependent increase of the Nczf promoteractivity. Expression of Nczf mRNA was detected in enteric neurons like that of Ncx. The amount of Nczf mRNA roughly correlated with that of the Ncx in enteric ganglia during embryonic development. Nczf localized in the nucleus and functioned as a transcriptional repressor. The consensus binding sequence of core nucleotides contains (A/T/C)CTTT(A/G)TTNT. In a gel mobility shift assay, the probe containing these sequences bound to the fusion protein. In silico analysis, these consensus sequences were found on regulatory regions of the endothelin receptor B and the microphthalmia-associated transcription factor genes, which are involved in neural crest development. Furthermore, overexpression of Nczf incultured neuroblastoma and fibroblast cell lines caused apoptotic cell death. These results suggest that Nczf functions as a sequence specific transcription repressor to regulate neural crest cell development.

Research Products

• [Journal Article] Pkd1 regulates immortalized proliferation of renal tubular epithelial cells through induction of P53 and activation of JNK. (2005)
  • Author(s): Nishio, S., et al.
  • Journal Title: J. Clin. Invest. 115 Pages: 910-918
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A role for c-Fos/Ap-1 in B lymphocyte terminal differentiation. (2005)
  • Author(s): Ohkubo, Y., et al.
  • Journal Title: J. Immunol. 174 Pages: 7703-7710
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Overexpression of Nd1-S, a variant form of new kelch family protein, perturbs the cell cycle progression of fibroblasts. (2005)
  • Author(s): Inoue, A., et al.
  • Journal Title: DNA Cell Biol. 24 Pages: 30-34
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Pkd1 regulates immortalized proliferation of renal tubular epithelial cells through induction ofP53 and activation of JNK. (2005)
  • Author(s): Nishio, S., Hatano, M., Nagata, M., Horie, S., Koike, T., Tokuhisa, T., Mochizuki, T.
  • Journal Title: J.Clin.Invest. 115 Pages: 910-918
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A role for c-Fos/AP-1 in B lymphocyte terminal differentiation. (2005)
  • Author(s): Ohkubo, Y., Arima, M., Arguni, E., Okada, S., Yamashita, K., Asari, S., Obata, S., Sakamoto, A., Hatano, M., O-Wang, J., Ebara, M., Saisho, H., Tokuhisa, T.
  • Journal Title: J.Immunol. 174 Pages: 7703-7710
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Overexpression of Nd1-S, a variant form of new kelch family protein, perturbs the cell cycle progression of fibroblasts. (2005)
  • Author(s): Inoue, A., Kang, M., Fujimura, L., Takamori, Y., Sasagawa, K., Itoh, H., Tokuhisa, T., Hatano, M.
  • Journal Title: DNA Cell Biol. 24 Pages: 30-34
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Abnormal erythroid differentiation in neonatal Bcl6-deficient mice. (2005)
  • Author(s): Asari, S., et al.
  • Journal Title: Exp.Hematol. 33 Pages: 26-34
• [Journal Article] Pkdl regulates immortalized proliferation of renal tubular epithelial cells through induction of P53 and activation of JNK. (2005)
  • Author(s): Nishio, S., et al.
  • Journal Title: J.Clin.Invest. 115 Pages: 910-918
• [Journal Article] Transcriptional activation of survivin through the NF-kappaB pathway by human T-cell leukemia virus type I tax. (2005)
  • Author(s): Kawakami, H., et al.
  • Journal Title: Int.J.Cancer 115 Pages: 967-974
• [Journal Article] A role for c-Fos/AP-1 in B lymphocyte terminal differentiation. (2005)
  • Author(s): Ohkubo, Y., et al.
  • Journal Title: J.Immunol. 174 Pages: 7703-7710
• [Journal Article] The pro-atherogenic cytokine interleukin-18 induces CXCL16 expression in rat aortic smooth muscle cells via MyD88, IRAK, TRAF6, c-Src, PI3K, Akt, JNK, and AP-1 signaling (2005)
  • Author(s): Chandrasekar, B., et al.
  • Journal Title: J.Biol.Chem. 280 Pages: 26263-26277
• [Journal Article] Overexpression of Nd1-S, a variant form of newkelch family protein, perturbs the cell cycle progression of fibroblasts. (2005)
  • Author(s): Inoue, A., et al.
  • Journal Title: DNA Cell Biol. 24 Pages: 30-34
• [Journal Article] Pkd1 regulates immortalized proliferation of renal tubular epithelial cells through p53 induction and JNK activation (2005)
  • Author(s): Nishio et al.
  • Journal Title: J Clin Invest. (In press)
• [Journal Article] Overexpression of Nd1-S, a variant form of new kelch family protein, perturbs the cell cycle progression of fibroblasts (2005)
  • Author(s): Inoue et al.
  • Journal Title: DNA and Cell Biol. 24・1 Pages: 30-34
• [Journal Article] Transcriptional activation of survivin through the NF-kappaB pathway by human T-cell leukemia virus type I tax (2005)
  • Author(s): Kawakami et al.
  • Journal Title: Int J Cancer (In press)
• [Journal Article] Acetylcholine-related bowel dysmotility in homozygous mutant NCX/HOX11L.1-deficient (NCX-/-) mice-evidence that acetylcholine is implicated in causing intestinal neuronal dysplasia. (2004)
  • Author(s): Yanai, T., et al.
  • Journal Title: J. Pediatr. Surg. 38 Pages: 927-930
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Protective role of Nd1 in doxorubicin-induced cardiotoxicity. (2004)
  • Author(s): Fujimura, L., et al.
  • Journal Title: Cardiovasc. Res. 64 Pages: 315-321
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Role of autophagy during the early neonatal starvation period. (2004)
  • Author(s): Kuma, A., et al.
  • Journal Title: Nature 432 Pages: 1032-1036
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Acetylcholine-related bowel dysmotility in homozygous mutant NCX/HOX11L.1-deficient (NCX-/-) mice-evidence that acetylcholine is implicated in causing intestinal neuronal dysplasia. (2004)
  • Author(s): Yanai T, Kobayashi H, Yamataka A, Lane GJ, Miyano T, Hayakawa T, Satoh K, Kase Y, Hatano M.
  • Journal Title: J Pediatr Surg. 38 Pages: 927-930
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Protective role of Nd1 in doxorubicin-induced cardiotoxicity. (2004)
  • Author(s): Fujimura, L., Matsudo, Y., Kang, M., Takamori, Y., Tokuhisa, T., Hatano, M.
  • Journal Title: Cardiovasc.Res. 64 Pages: 315-321
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Role of autophagy during the early neonatal starvation period. (2004)
  • Author(s): Kuma, A., Hatano, M., Matsui, M., Yamamoto, A., Nakaya, H., Yoshimori, T., Ohsumi, Y., Tokuhisa, T., Mizushima, N.
  • Journal Title: Nature 432 Pages: 1032-1036
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Acetylcholine-related bowel dysmotility in homozygous mutant NCX/HOX11L1-deficient (NCX-/-) mice-evidence that acetylcholine is implicated in causing intestinal neuronal dysplasia (2004)
  • Author(s): Yanai et al.
  • Journal Title: J Pediatr Surg. 39・6 Pages: 927-930
• [Journal Article] The role of autophagy during the early neonatal starvation period (2004)
  • Author(s): Kuma et al.
  • Journal Title: Nature 432・7020 Pages: 1032-1036
• [Journal Article] Protective role of Nd1 in doxorubicin-induced cardiotoxicity (2004)
  • Author(s): Fujimura et al.
  • Journal Title: Cardiovasc Res. 64・2 Pages: 315-321
• [Book] Strategies for disease control. (2005)
  • Author(s): Arguni, E., et al.
  • Total Pages: 216
  • Publisher: Elsevier
  • Description: 「研究成果報告書概要(和文)」より