title: CCN3 protein participates in bone regeneration as an inhibitory factor.
abstract: CCN3, a member of the CCN protein family, inhibits osteoblast differentiation in vitro. However, the role of CCN3 in bone regeneration has not been well elucidated. In this study, we investigated the role of CCN3 in bone regeneration. We identified the Ccn3 gene by microarray analysis as a highly expressed gene at the early phase of bone regeneration in a mouse bone regeneration model. We confirmed the up-regulation of Ccn3 at the early phase of bone regeneration by RT-PCR, Western blot, and immunofluorescence analyses. Ccn3 transgenic mice, in which Ccn3 expression was driven by 2.3-kb Col1a1 promoter, showed osteopenia compared with wild-type mice, but Ccn3 knock-out mice showed no skeletal changes compared with wild-type mice. We analyzed the bone regeneration process in Ccn3 transgenic mice and Ccn3 knock-out mice by microcomputed tomography and histological analyses. Bone regeneration in Ccn3 knock-out mice was accelerated compared with that in wild-type mice. The mRNA expression levels of osteoblast-related genes (Runx2, Sp7, Col1a1, Alpl, and Bglap) in Ccn3 knock-out mice were up-regulated earlier than those in wild-type mice, as demonstrated by RT-PCR. Bone regeneration in Ccn3 transgenic mice showed no significant changes compared with that in wild-type mice. Phosphorylation of Smad1/5 was highly up-regulated at bone regeneration sites in Ccn3 KO mice compared with wild-type mice. These results indicate that CCN3 is up-regulated in the early phase of bone regeneration and acts as a negative regulator for bone regeneration. This study may contribute to the development of new strategies for bone regeneration therapy.
authors: Section of Oral Pathology, International Research Center for Molecular Science in Tooth and Bone Diseases, Tokyo Medical and Dental University, Tokyo 113-8549, Japan.
authors: Yuki
authors: Y
authors: Matsushita
authors: Kei
authors: K
authors: Sakamoto
authors: Yoshihiro
authors: Y
authors: Tamamura
authors: Yasuaki
authors: Y
authors: Shibata
authors: Tokutaro
authors: T
authors: Minamizato
authors: Tasuku
authors: T
authors: Kihara
authors: Masako
authors: M
authors: Ito
authors: Ken-ichi
authors: K
authors: Katsube
authors: Shuichi
authors: S
authors: Hiraoka
authors: Haruhiko
authors: H
authors: Koseki
authors: Kiyoshi
authors: K
authors: Harada
authors: Akira
authors: A
authors: Yamaguchi
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D001861:Bone Regeneration; D024042:Collagen Type I / Q000096:biosynthesis / Q000235:genetics; D000090602:Collagen Type I, alpha 1 Chain; D050659:Core Binding Factor Alpha 1 Subunit / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D055514:Nephroblastoma Overexpressed Protein / Q000096:biosynthesis* / Q000235:genetics; D010006:Osteoblasts / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D051898:Smad1 Protein / Q000096:biosynthesis / Q000235:genetics; D051902:Smad5 Protein / Q000096:biosynthesis / Q000235:genetics; D000076182:Sp7 Transcription Factor; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D015854:Up-Regulation; D055114:X-Ray Microtomography
chemical_list: D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D050659:Core Binding Factor Alpha 1 Subunit; D055514:Nephroblastoma Overexpressed Protein; C493716:Runx2 protein, mouse; D051898:Smad1 Protein; C497401:Smad1 protein, mouse; D051902:Smad5 Protein; C497798:Smad5 protein, mouse; D000076182:Sp7 Transcription Factor; C479466:Sp7 protein, mouse; D014157:Transcription Factors; D000469:Alkaline Phosphatase
keywords: Bone; Bone Morphogenetic Protein (BMP); CCN3; Knockout Mice; Osteoblasts; Regeneration; Transgenic Mice
references: J Cell Biol. 1994 Dec;127(6 Pt 1):1755-66
references: 7798324
references: BMC Med. 2011 May 31;9:66
references: 21627784
references: Int Orthop. 2009 Feb;33(1):281-8
references: 19023570
references: Arterioscler Thromb Vasc Biol. 2000 Aug;20(8):1912-9
references: 10938011
references: Bone. 2010 Dec;47(6):1006-12
references: 20807599
references: BMC Dev Biol. 2008 Feb 20;8:18
references: 18289368
references: Arterioscler Thromb Vasc Biol. 2010 Apr;30(4):675-82
references: 20139355
references: Biochem Biophys Res Commun. 2000 Jul 5;273(2):630-6
references: 10873656
references: J Bone Miner Res. 2005 Oct;20(10):1828-36
references: 16160740
references: J Clin Invest. 1998 Dec 15;102(12):2106-14
references: 9854046
references: Am J Pathol. 1998 Jun;152(6):1563-75
references: 9626060
references: Science. 2007 Apr 27;316(5824):590-3
references: 17463287
references: Dev Dyn. 2000 Nov;219(3):417-25
references: 11066097
references: Lancet. 2004 Jan 3;363(9402):62-4
references: 14723997
references: Mol Cell Biol. 1993 Sep;13(9):5168-74
references: 8355676
references: J Virol. 1989 Apr;63(4):1803-7
references: 2564440
references: J Orthop Res. 2009 Dec;27(12):1652-8
references: 19489046
references: Endocrinology. 2010 Jan;151(1):221-33
references: 19934377
references: J Bone Miner Res. 2001 May;16(5):876-84
references: 11341332
references: Bone. 2012 May;50(5):1081-91
references: 22387305
references: J Cell Biol. 2001 Oct 1;155(1):157-66
references: 11581292
references: Biochem Biophys Res Commun. 2008 Apr 11;368(3):808-14
references: 18275847
references: J Orthop Res. 1995 May;13(3):357-67
references: 7602397
references: J Cell Biol. 1991 May;113(3):681-7
references: 1849907
references: FEBS Lett. 2011 Oct 3;585(19):3033-40
references: 21871891
references: J Bone Miner Res. 1987 Dec;2(6):595-610
references: 3455637
references: J Bone Miner Metab. 2006;24(6):425-33
references: 17072733
references: Am J Pathol. 2011 May;178(5):2377-88
references: 21514448
references: J Biol Chem. 2005 Mar 4;280(9):8229-37
references: 15611078
references: Calcif Tissue Int. 2010 Jan;86(1):72-81
references: 19953233
references: J Cell Commun Signal. 2009 Jun;3(2):135-45
references: 19626464
references: J Cell Commun Signal. 2007 Dec;1(3-4):159-64
references: 18568428
references: Bone. 2000 Jul;27(1):81-90
references: 10865213
references: Endocr Rev. 2000 Aug;21(4):393-411
references: 10950158
references: C R Acad Sci III. 1998 Nov;321(11):883-92
references: 9879467
references: Bone. 2008 Jun;42(6):1144-53
references: 18372226
references: J Clin Endocrinol Metab. 1999 Mar;84(3):1096-103
references: 10084601
references: J Cell Commun Signal. 2012 Jun;6(2):73-85
references: 22427255
references: Brain Res Gene Expr Patterns. 2001 Aug;1(1):61-5
references: 15018819
references: Bone. 2011 Nov;49(5):975-89
references: 21763478
references: Endocrinology. 2000 Dec;141(12):4558-63
references: 11108268
references: Biochem Biophys Res Commun. 2007 Mar 9;354(2):567-73
references: 17250806
references: Virchows Arch. 2008 Jan;452(1):33-9
references: 18066593
references: J Anat. 2010 May;216(5):625-30
references: 20298438
references: J Biol Chem. 2002 Aug 16;277(33):30177-82
references: 12055193
references: J Biol Chem. 2007 Jul 6;282(27):19762-72
references: 17500060
references: J Biol Chem. 2002 Aug 16;277(33):29399-405
references: 12050162
references: J Bone Miner Res. 2005 Dec;20(12):2213-23
references: 16294274
references: Bone. 2012 Jul;51(1):59-68
references: 22521262