PMID- 32558157 OWN - NLM STAT- MEDLINE DCOM- 20210427 LR - 20220531 IS - 1469-3178 (Electronic) IS - 1469-221X (Print) IS - 1469-221X (Linking) VI - 21 IP - 8 DP - 2020 Aug 5 TI - CTGF/CCN2 facilitates LRP4-mediated formation of the embryonic neuromuscular junction. PG - e48462 LID - 10.15252/embr.201948462 [doi] LID - e48462 AB - At the neuromuscular junction (NMJ), lipoprotein-related receptor 4 (LRP4) mediates agrin-induced MuSK phosphorylation that leads to clustering of acetylcholine receptors (AChRs) in the postsynaptic region of the skeletal muscle. Additionally, the ectodomain of LRP4 is necessary for differentiation of the presynaptic nerve terminal. However, the molecules regulating LRP4 have not been fully elucidated yet. Here, we show that the CT domain of connective tissue growth factor (CTGF/CCN2) directly binds to the third beta-propeller domain of LRP4. CTGF/CCN2 enhances the binding of LRP4 to MuSK and facilitates the localization of LRP4 on the plasma membrane. CTGF/CCN2 enhances agrin-induced MuSK phosphorylation and AChR clustering in cultured myotubes. Ctgf-deficient mouse embryos (Ctgf(-/-) ) have small AChR clusters and abnormal dispersion of synaptic vesicles along the motor axon. Ultrastructurally, the presynaptic nerve terminals have reduced numbers of active zones and mitochondria. Functionally, Ctgf(-/-) embryos exhibit impaired NMJ signal transmission. These results indicate that CTGF/CCN2 interacts with LRP4 to facilitate clustering of AChRs at the motor endplate and the maturation of the nerve terminal. CI - (c) 2020 The Authors. FAU - Ohkawara, Bisei AU - Ohkawara B AUID- ORCID: 0000-0002-1987-458X AD - Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan. FAU - Kobayakawa, Akinori AU - Kobayakawa A AD - Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan. AD - Department of Orthopedic Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan. FAU - Kanbara, Shunsuke AU - Kanbara S AD - Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan. AD - Department of Orthopedic Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan. FAU - Hattori, Takako AU - Hattori T AD - Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan. FAU - Kubota, Satoshi AU - Kubota S AD - Department of Biochemistry and Molecular Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan. FAU - Ito, Mikako AU - Ito M AD - Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan. FAU - Masuda, Akio AU - Masuda A AUID- ORCID: 0000-0002-6437-2853 AD - Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan. FAU - Takigawa, Masaharu AU - Takigawa M AD - Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Dental School, Okayama, Japan. FAU - Lyons, Karen M AU - Lyons KM AD - Department of Orthopedic Surgery, UCLA, Los Angeles, CA, USA. FAU - Ishiguro, Naoki AU - Ishiguro N AD - Department of Orthopedic Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan. FAU - Ohno, Kinji AU - Ohno K AD - Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan. LA - eng GR - JP18K06483/Grants-in-Aids from the Japan Society for the Promotion of Science/International GR - JP17K07094/Grants-in-Aids from the Japan Society for the Promotion of Science/International GR - JP18K06058/Grants-in-Aids from the Japan Society for the Promotion of Science/International GR - JP15H05014/Grants-in-Aids from the Japan Society for the Promotion of Science/International GR - JP17K19757/Grants-in-Aids from the Japan Society for the Promotion of Science/International GR - JP19H03817/Grants-in-Aids from the Japan Society for the Promotion of Science/International GR - JP15H05015/Grants-in-Aids from the Japan Society for the Promotion of Science/International GR - H29-Nanchi-Ippan-030/The Ministry of Health, Labour and Welfare of Japan/International GR - JP19gm1010002/The Japan Agency for Medical Research and Development/International GR - JP19ek0109230/The Japan Agency for Medical Research and Development/International GR - JP19ek0109281/The Japan Agency for Medical Research and Development/International GR - JP19bm0804005/The Japan Agency for Medical Research and Development/International GR - 29-4/Intramural Research Grant for Neurological and Psychiatric Disorders of NCNP/International PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20200617 PL - England TA - EMBO Rep JT - EMBO reports JID - 100963049 RN - 0 (Agrin) RN - 0 (LDL-Receptor Related Proteins) RN - 0 (Lrp4 protein, mouse) RN - 139568-91-5 (Connective Tissue Growth Factor) SB - IM MH - Agrin/genetics/metabolism MH - Animals MH - *Connective Tissue Growth Factor/genetics/metabolism MH - *LDL-Receptor Related Proteins/genetics/metabolism MH - Mice MH - Neuromuscular Junction/metabolism MH - Phosphorylation PMC - PMC7403661 OTO - NOTNLM OT - acetylcholine receptor OT - cellular communication network factor 2 OT - connective tissue growth factor OT - low-density lipoprotein-related receptor 4 OT - neuromuscular junction COIS- The authors declare that they have no conflict of interest. EDAT- 2020/06/20 06:00 MHDA- 2021/04/28 06:00 PMCR- 2021/08/05 CRDT- 2020/06/20 06:00 PHST- 2019/05/17 00:00 [received] PHST- 2020/05/20 00:00 [revised] PHST- 2020/05/27 00:00 [accepted] PHST- 2020/06/20 06:00 [pubmed] PHST- 2021/04/28 06:00 [medline] PHST- 2020/06/20 06:00 [entrez] PHST- 2021/08/05 00:00 [pmc-release] AID - EMBR201948462 [pii] AID - 10.15252/embr.201948462 [doi] PST - ppublish SO - EMBO Rep. 2020 Aug 5;21(8):e48462. doi: 10.15252/embr.201948462. Epub 2020 Jun 17.