Identification of specific splicing regulation at neuromuscular junctions

• Project/Area Number: 24500469
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurophysiology and muscle physiology
• Research Institution: Nagoya University
• Principal Investigator: Masuda Akio 名古屋大学, 医学(系)研究科(研究院), 准教授 (10343203)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 神経筋接合部 / スプライシング / 筋分化 / RNA-seq / Agrin / レーザーマイクロダイセクション / RNA代謝

Outline of Final Research Achievements

To analyze specific regulation of RNA processing in neuromucular junctions, we harvested RNA from neuromuscular junctions in mouse skeletal muscles with laser microdissection, and performed microarray and RNA-seq analysis. Our analysis revealed that alternative splicing was distinctly regulated at neuromuscular junctions.
We also examined C2C12 cmyotubes stimulated with Agrin/Neuregulin, and found that SR proteins were deeply involved in the specific regulation of mRNA splicing at neuromuscular junctions.

Research Products

• [Journal Article] Collagen Q and anti-MuSK autoantibody competitively suppress agrin/LRP4/MuSK signaling. (2015)
  • Author(s): Otsuka K, Ito M, Ohkawara B, Masuda A, Kawakami Y, Sahashi K, Nishida H, Mabuchi N, Takano A, Engel AG, Ohno K.
  • Journal Title: Sci Rep. Volume: 5 Issue: 1 Pages: 13928-13928
  • DOI: 10.1038/srep13928
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] SRSF1 and hnRNP H antagonistically regulate splicing of COLQ exon 16 in a congenital myasthenic syndrome. (2015)
  • Author(s): Rahman MA, Azuma Y, Nasrin F, Takeda J, Nazim M, Bin Ahsan K, Akio Masuda, Engel AG, Ohno K.
  • Journal Title: Sci Rep. Volume: 5 Issue: 1 Pages: 13208-13208
  • DOI: 10.1038/srep13208
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Position-specific binding of FUS to nascent RNA regulates mRNA length. (2015)
  • Author(s): Masuda A, Takeda J, Okuno T, Okamoto T, Ohkawara B, Ito M, Ishigaki S, Sobue G, Ohno K.
  • Journal Title: Genes Dev. Volume: 29(10) Issue: 10 Pages: 1045-57
  • DOI: 10.1101/gad.255737.114
  • Peer Reviewed / Open Access
• [Journal Article] Congenital myasthenic syndrome in Japan: ethnically unique mutations in muscle nicotinic acetylcholine receptor subunits (2015)
  • Author(s): Azuma Y, Nakata T, Tanaka M, Shen X M, Ito M, Iwata S, Okuno T, Nomura Y, Ando N, Ishigaki K, Ohkawara B, Masuda A, Natsume J, Kojima S, Sokabe M, Ohno K
  • Journal Title: Neuromuscul Disord Volume: 25 Issue: 2 Pages: 60-69
  • DOI: 10.1210/en.2014-1914
  • Peer Reviewed
• [Journal Article] A missense mutation in domain III in HSPG2 in Schwartz-Jampel syndrome compromises secretion of perlecan into the extracellular space. (2015)
  • Author(s): Iwata S, Ito M, Nakata T, Noguchi Y, Okuno T, Ohkawara B, Masuda A, Goto T, Adachi M, Osaka H, Nonaka R, Arikawa-Hirasawa E, Ohno K.
  • Journal Title: Neuromuscul Disord. Volume: 25(8) Issue: 8 Pages: 667-71
  • DOI: 10.1016/j.nmd.2015.05.002
  • Peer Reviewed
• [Journal Article] Meclozine promotes longitudinal skeletal growth in transgenic mice with achondroplasia carrying a gain-of-function mutation in the FGFR3 gene (2015)
  • Author(s): Matsushita M, Hasegawa S, Kitoh H, Mori K, Ohkawara B, Yasoda A, Masuda A, Ishiguro N, Ohno K,
  • Journal Title: Endocrinology Volume: 156 Issue: 1 Pages: 584-554
  • DOI: 10.1016/j.fob.2014.11.007
  • Peer Reviewed / Open Access
• [Journal Article] HnRNP C, YB-1 and hnRNP L coordinately enhance skipping of human MUSK exon 10 to generate a Wnt-insensitive MuSK isoform (2014)
  • Author(s): Nasrin F, Rahman MA, Masuda A, Ohe K, Takeda J, Ohno K.
  • Journal Title: Sci Rep Volume: 4 Issue: 1 Pages: 6841-6841
  • DOI: 10.1038/srep06841
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] LRP4 induces extracellular matrix productions and facilitates chondrocyte differentiation (2014)
  • Author(s): Asai N, Ohkawara B, Ito M, Masuda A, Ishiguro N, Ohno K
  • Journal Title: Biochem Biophys Res Commun. Volume: 451 Issue: 2 Pages: 302-307
  • DOI: 10.1016/j.bbrc.2014.07.125
  • Peer Reviewed / Open Access
• [Journal Article] Verapamil protects against cartilage degradation in osteoarthritis by inhibiting Wnt/β-catenin signaling. (2014)
  • Author(s): Takamatsu, A., Ohkawara, B., Ito, M., Masuda, A., Sakai, T., Ishiguro, N. and Ohno, K.
  • Journal Title: PLOS ONE Volume: 9(3) Issue: 3 Pages: e92699-e92699
  • DOI: 10.1371/journal.pone.0092699
  • Peer Reviewed / Open Access
• [Journal Article] LRP4 third β-propeller domain mutations cause novel congenital myasthenia by compromising agrin-mediated MuSK signaling in a position-specific manner (2014)
  • Author(s): Ohkawara B, Cabrera-Serrano M, Nakata T, Milone M, Asai N, Ito K, Ito M, Masuda A, Ito Y, Engel AG, Ohno K
  • Journal Title: Hum Mol Genet. Volume: Apr 1;23(7) Issue: 7 Pages: 1856-68
  • DOI: 10.1093/hmg/ddt578
  • Peer Reviewed / Open Access
• [Journal Article] Meclozine facilitates proliferation and differentiation of chondrocytes by attenuating abnormally activated FGFR3 signaling in achondroplasia. (2013)
  • Author(s): Matsushita, M., Kitoh, H., Ohkawara, B., Mishima, K., Kaneko, H., Ito, M., Masuda, A., Ishiguro, N. and Ohno, K.
  • Journal Title: PLOS ONE Volume: 8(12) Issue: 12 Pages: 81569-81569
  • DOI: 10.1371/journal.pone.0081569
  • Peer Reviewed
• [Journal Article] The ALS/FTLD-related RNA-binding proteins TDP-43 and FUS have common downstream RNA targets in cortical neurons (2013)
  • Author(s): Honda D, Ishigaki S, Iguchi Y, Fujioka Y, Udagawa T, Masuda A, Ohno K, Katsuno M, Sobue G.
  • Journal Title: FEBS Open Bio. Volume: 20 Issue: 1 Pages: 1-10
  • DOI: 10.1016/j.fob.2013.11.001
  • Peer Reviewed
• [Journal Article] HnRNP L and hnRNP LL antagonistically modulate PTB-mediated splicing suppression of CHRNA1 pre-mRNA. (2013)
  • Author(s): Rahman, M. A., Masuda, A., Ohe, K., Ito, M. , Hutchinson, D. O., Mayeda, A., Engel, A. G. and Ohno, K.
  • Journal Title: Sci. Rep. Volume: 3 Issue: 1 Pages: 2931-2931
  • DOI: 10.1038/srep02931
  • Peer Reviewed
• [Journal Article] FUS-regulated region- and cell-type-specific transcriptome is associated with cell selectivity in ALS/FTLD. (2013)
  • Author(s): Fujioka Y, Ishigaki S, Masuda A, Iguchi Y, Udagawa T, Watanabe H, Katsuno M, Ohno K, Sobue G.
  • Journal Title: Sci Rep. Volume: 3 Issue: 1 Pages: 2388-2388
  • DOI: 10.1038/srep02388
  • Peer Reviewed
• [Journal Article] Mutations in the C-Terminal Domain of ColQ in Endplate Acetylcholinesterase Deficiency Compromise ColQ-MuSK Interaction. (2013)
  • Author(s): Nakata T, Ito M, Azuma Y, Otsuka K, Noguchi Y, Komaki H, Okumura A, Shiraishi K, Masuda A, Natsume J, Kojima S, Ohno K.
  • Journal Title: Human Mutation Volume: in press Issue: 7 Pages: 997-1004
  • DOI: 10.1002/humu.22325
  • Peer Reviewed
• [Journal Article] Clinically applicable antianginal agents suppress osteoblastic transformation of myogenic cells and heterotopic ossifications in mice (2013)
  • Author(s): Yamamoto R, Matsushita M, Kitoh H, Masuda A, Ito M, Katagiri T, Kawai T, Ishiguro N, Ohno K
  • Journal Title: J Bone Miner Metab Volume: 31 Pages: 26-33
  • DOI: 10.1007/s00774-012-0380-2
  • NAID: 10031168834
  • Peer Reviewed
• [Journal Article] Position-dependent FUS-RNA interactions regulate alternative splicing events and transcriptions. (2012)
  • Author(s): Ishigaki S, Masuda A, Fujioka Y, Iguchi Y, Katsuno M, Shibata A, Urano F, Sobue G, Ohno K
  • Journal Title: Sci Rep Volume: 2 Issue: 1 Pages: 529-529
  • DOI: 10.1038/srep00529
  • Peer Reviewed
• [Journal Article] Four parameters increase the sensitivity and specificity of the exon array analysis and disclose twenty-five novel aberrantly spliced exons in myotonic dystrophy. (2012)
  • Author(s): Yamashita Y, Matsuura T, Shinmi J, Amakusa Y, Masuda A, Ito M, Kinoshita M, Furuya H, Abe K, Ibi T, Sahashi K, Ohno K.
  • Journal Title: Journal of Human Genetics Volume: 57 Issue: 6 Pages: 368-374
  • DOI: 10.1038/jhg.2012.37
  • Peer Reviewed
• [Journal Article] Protein-anchoring strategy for delivering acetylcholinesterase to the neuromuscular junction (2012)
  • Author(s): Ito M, Suzuki Y, Okada T, Fukudome T, Yoshimura T, Masuda A, Takeda S, Krejci E, Ohno K
  • Journal Title: Mol Ther Volume: (印刷中) Issue: 7 Pages: 1384-1392
  • DOI: 10.1038/mt.2012.34
  • Peer Reviewed
• [Presentation] 神経変性疾患関連RNA結合タンパクFUSによる早期転写終結機構の解明 (2014)
  • Author(s): Akio Masuda, Junichi Takeda, Tatsuya Okuno, Kinji Ohno
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜(神奈川県横浜市)
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] Genome-wide analysis reveals relationship between RNA-binding sites of FUS and transcriptional regulation (2013)
  • Author(s): Akio Masuda, Tatsuya Okuno, Takaaki Okamoto, Kinji Ohno
  • Organizer: 8th Brain Research Conference: RNA Metabolism in Neurological Disease
  • Place of Presentation: San Diego, USA
• [Presentation] FUS regulates transcription termination through binding to nascent RNA (2013)
  • Author(s): Akio Masuda, Tatsuya Okuno, Takaaki Okamoto, Kinji Ohno
  • Organizer: Society for Neuroscience
  • Place of Presentation: San Diego, USA
• [Presentation] Masuda A, Andersen HS, Doktor TK, Okamoto T, Ito M, Andresen BS, Ohno K. (2012)
  • Author(s): Global analysis of RNA-binding sites of CUGBP1 and MBNL1 reveal their preferential binding to 3’ UTRs to regulate mRNA stability
  • Organizer: The 11th Annual Scientific Meeting of Asian and Oceanian Myology Center
  • Place of Presentation: Kyoto
• [Presentation] Akio Masuda, Shinsuke Ishigaki, Yusuke Fujioka, Yohei Iguchi, Masahisa Katsuno, Akihide Shibata, Fumihiko Urano, Gen Sobue, Kinji Ohno (2012)
  • Author(s): Global mapping of FUS-binding sites and global profiling of FUS-mediated RNA metabolisms in the mouse brain
  • Organizer: Annual meeting of American Society of Human Genetics
  • Place of Presentation: San Francisco, USA
• [Remarks] 先端応用医学部門・神経遺伝情報分野
  • URL: http://www.med.nagoya-u.ac.jp/neurogenetics
• [Remarks] 先端応用医学部門 神経遺伝情報学分野
  • URL: http://www.med.nagoya-u.ac.jp/neurogenetics/
• [Remarks] 先端応用医学部門 神経遺伝情報分野
  • URL: http://www.med.nagoya-u.ac.jp/neurogenetics/