Yuichiro Enoki1, Tsuyoshi Sato2, Shoichiro Kokabu1,3, Naoki Hayashi1, Takanori Iwata4, Masayuki Yamato4, Michihiko Usui5, Masahito Matsumoto6, Taketo Tomoda1, Wataru Ariyoshi7, Tatsuji Nishihara7, Tetsuya Yoda1. 1. Department of Oral and Maxillofacial Surgery, Saitama Medical University, Saitama, Japan. 2. Department of Oral and Maxillofacial Surgery, Saitama Medical University, Saitama, Japan tsato@saitama-med.ac.jp. 3. Division of Molecular Signaling and Biochemistry, Department of Health Promotion and Kyushu Dental University, Fukuoka, Japan. 4. Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, Tokyo, Japan. 5. Division of Periodontology, Department of Cardiology and Periodontology, Kyushu Dental University, Fukuoka, Japan. 6. Division of Functional Genomics and Systems Medicine, Research Center for Genomic Medicine, Saitama Medical University, Saitama, Japan. 7. Division of Infections and Molecular Biology, Department of Health Promotion, Kyushu Dental University, Fukuoka, Japan.
Abstract
BACKGROUND/AIM: While netrin-4 plays a vital role in the vascular system, the role of netrin-1 in osteoblast differentiation is not well understood. In this study we explored whether netrin-4 has functional roles in osteoblasts. MATERIALS AND METHODS: Quantitative reverse-transcriptase polymerase chain reaction (PCR), RNA interference, the generation of plasmids, transfections, measurement of alkaline phosphatase activity, a mineralization assay, a migration assay and a cell proliferation assay were performed. RESULTS: Netrin-4 expression was up-regulated during osteoblast differentiation and an RNA interference experiment showed that small interfering RNA used to silence netrin-4 inhibited osteoblast differentiation. Recombinant mouse netrin-4 promoted alkaline phosphatase (ALP) activity of osteoblasts and enhancement of calcium deposits. Moreover, we constructed a vector containing the netrin-4 gene on the basis of the plasmid pcDNA3.1/V5-His. Overexpression of netrin-4 enhanced differentiation of osteoblasts. Finally, recombinant mouse netrin-4 promoted cell migration of osteoblasts. CONCLUSION: Netrin-4 promotes differentiation and migration of osteoblasts. Copyright
BACKGROUND/AIM: While netrin-4 plays a vital role in the vascular system, the role of netrin-1 in osteoblast differentiation is not well understood. In this study we explored whether netrin-4 has functional roles in osteoblasts. MATERIALS AND METHODS: Quantitative reverse-transcriptase polymerase chain reaction (PCR), RNA interference, the generation of plasmids, transfections, measurement of alkaline phosphatase activity, a mineralization assay, a migration assay and a cell proliferation assay were performed. RESULTS:Netrin-4 expression was up-regulated during osteoblast differentiation and an RNA interference experiment showed that small interfering RNA used to silence netrin-4 inhibited osteoblast differentiation. Recombinant mousenetrin-4 promoted alkaline phosphatase (ALP) activity of osteoblasts and enhancement of calcium deposits. Moreover, we constructed a vector containing the netrin-4 gene on the basis of the plasmid pcDNA3.1/V5-His. Overexpression of netrin-4 enhanced differentiation of osteoblasts. Finally, recombinant mousenetrin-4 promoted cell migration of osteoblasts. CONCLUSION:Netrin-4 promotes differentiation and migration of osteoblasts. Copyright
Authors: N Udagawa; N Takahashi; E Jimi; K Matsuzaki; T Tsurukai; K Itoh; N Nakagawa; H Yasuda; M Goto; E Tsuda; K Higashio; M T Gillespie; T J Martin; T Suda Journal: Bone Date: 1999-11 Impact factor: 4.398
Authors: Mayra Yebra; Giuseppe R Diaferia; Anthony M P Montgomery; Thomas Kaido; William J Brunken; Manuel Koch; Gary Hardiman; Laura Crisa; Vincenzo Cirulli Journal: PLoS One Date: 2011-07-29 Impact factor: 3.240
Authors: Masahiko Okubo; K O Ito; Fumie Yamazaki; Yosuke Mizuno; Yuta Isozaki; Seiji Asoda; Michihiko Usui; Tsuyoshi Sato Journal: In Vivo Date: 2022 Jul-Aug Impact factor: 2.406