Luciferase transgenic mice are a very promising tool for noninvasive, quantitative, and longitudinal evaluation of gene expression. The aim of this study was to validate the Col(I)-Luc transgenic mouse model in which the luciferase gene is driven by bone-specific regulatory elements from the mouse collagen α1(I) gene for bioluminescent imaging of bone development and remodeling. We observed strong luciferase activity in skeletal tissues of Col(I)-Luc mice, and observed that the light intensity declined with postnatal bone development. Luciferase activity was enhanced in a tail bone repair model and we were able to monitor the process of ectopic bone formation induced by recombinant human bone morphogenetic protein 2 using bioluminescent imaging. We conclude that Col(I)-Luc transgenic mice can be applied in the field of bone tissue engineering for monitoring bone repair processes and for investigating osteoinductive molecules or scaffolds.
HartmanE.H., PikkemaatJ.A., VehofJ.W., HeerschapA., JansenJ.A., SpauwenP.H.In vivo magnetic resonance imaging explorative study of ectopic bone formation in the rat. Tissue Eng, 8:1029. 2002.
4.
BremerC., WeisslederR.In vivo imaging of gene expression. Acad Radiol, 8:15. 2001.
5.
RayP., DeA., MinJ.J., TsienR.Y., GambhirS.S.Imaging tri-fusion multimodality reporter gene expression in living subjects. Cancer Res, 64:1323. 2004.
6.
NtziachristosV., RipollJ., WangL.V., WeisslederR.Looking and listening to light: the evolution of whole-body photonic imaging. Nat Biotechnol, 23:313. 2005.
7.
HerschmanH.R.Molecular imaging: looking at problems, seeing solutions. Science, 302:605. 2003.
8.
de BoerJ., van BlitterswijkC., LowikC.Bioluminescent imaging: emerging technology for non-invasive imaging of bone tissue engineering. Biomaterials, 27:1851. 2006.
9.
MaggiA., OttobriniL., BiserniA., LucignaniG., CianaP.Techniques: reporter mice—a new way to look at drug action. Trends Pharmacol Sci, 25:337. 2004.
10.
BlumJ.S., TemenoffJ.S., ParkH., JansenJ.A., MikosA.G., BarryM.A.Development and characterization of enhanced green fluorescent protein and luciferase expressing cell line for non-destructive evaluation of tissue engineering constructs. Biomaterials, 25:5809. 2004.
11.
OlivoC., AlblasJ., VerweijV., Van ZonneveldA.J., DhertW.J., MartensA.C.In vivo bioluminescence imaging study to monitor ectopic bone formation by luciferase gene marked mesenchymal stem cells. J Orthop Res, 26:901. 2008.
12.
KumarS., MahendraG., PonnazhaganS.Determination of osteoprogenitor-specific promoter activity in mouse mesenchymal stem cells by recombinant adeno-associated virus transduction. Biochim Biophys Acta, 1731:95. 2005.
13.
MonteiroR.M., de Sousa LopesS.M., KorchynskyiO., ten DijkeP., MummeryC.L.Spatio-temporal activation of Smad1 and Smad5 in vivo: monitoring transcriptional activity of Smad proteins. J Cell Sci, 117:4653. 2004.
14.
LiuJ., BarradasA., FernandesH., JanssenF., PapenburgB., StamatialisD., MartensA.C., van BlitterswijkC.A., de BoerJ.In vitro and in vivo bioluminescence imaging of hypoxia in tissue engineered grafts. Tissue Eng Part C Methods, 2009Epub ahead of print.
15.
IrisB., ZilbermanY., ZeiraE., GalunE., HonigmanA., TurgemanG., ClemensT., GazitZ., GazitD.Molecular imaging of the skeleton: quantitative real-time bioluminescence monitoring gene expression in bone repair and development. J Bone Miner Res, 18:570. 2003.
16.
GafniY., PelledG., ZilbermanY., TurgemanG., ApparaillyF., YotvatH., GalunE., GazitZ., JorgensenC., GazitD.Gene therapy platform for bone regeneration using an exogenously regulated, AAV-2-based gene expression system. Mol Ther, 9:587. 2004.
17.
BogdanovicZ., BedalovA., KrebsbachP.H., PavlinD., WoodyC.O., ClarkS.H., ThomasH.F., RoweD.W., KreamB.E., LichtlerA.C.Upstream regulatory elements necessary for expression of the rat COL1A1 promoter in transgenic mice. J Bone Miner Res, 9:285. 1994.
18.
KalajzicI., KalajzicZ., KaliternaM., GronowiczG., ClarkS.H., LichtlerA.C., RoweD.Use of type I collagen green fluorescent protein transgenes to identify subpopulations of cells at different stages of the osteoblast lineage. J Bone Miner Res, 17:15. 2002.
19.
BobanI., JacquinC., PriorK., Barisic-DujmovicT., MayeP., ClarkS.H., AguilaH.L.The 3.6 kb DNA fragment from the rat Col1a1 gene promoter drives the expression of genes in both osteoblast and osteoclast lineage cells. Bone, 39:1302. 2006.
RossertJ., EberspaecherH., de CrombruggheB.Separate cis-acting DNA elements of the mouse pro-alpha 1(I) collagen promoter direct expression of reporter genes to different type I collagen-producing cells in transgenic mice. J Cell Biol, 129:1421. 1995.
22.
RossertJ.A., ChenS.S., EberspaecherH., SmithC.N., de CrombruggheB.Identification of a minimal sequence of the mouse pro-alpha 1(I) collagen promoter that confers high-level osteoblast expression in transgenic mice and that binds a protein selectively present in osteoblasts. Proc Natl Acad Sci USA, 93:1027. 1996.
23.
OpasE.E., GentileM.A., RossertJ.A., de CrombruggheB., RodanG.A., SchmidtA.Parathyroid hormone and prostaglandin E2 preferentially increase luciferase levels in bone of mice harboring a luciferase transgene controlled by elements of the pro-alpha1(I) collagen promoter. Bone, 26:27. 2000.
24.
LiX., GuW., MasindeG., Hamilton-UllandM., RundleC.H., MohanS., BaylinkD.J.Genetic variation in bone-regenerative capacity among inbred strains of mice. Bone, 29:134. 2001.
25.
YamamotoM., FisherJ.E., GentileM., SeedorJ.G., LeuC.T., RodanS.B., RodanG.A.The integrin ligand echistatin prevents bone loss in ovariectomized mice and rats. Endocrinology, 139:1411. 1998.
26.
TeixeiraS., FernandesH., LeusinkA., van BlitterswijkC., FerrazM.P., MonteiroF.J., de BoerJ.In vivo evaluation of highly macroporous ceramic scaffolds for bone tissue engineering. J Biomed Mater Res A, 93:567. 2010.
27.
TroyT., Jekic-McMullenD., SambucettiL., RiceB.Quantitative comparison of the sensitivity of detection of fluorescent and bioluminescent reporters in animal models. Mol Imaging, 3:9. 2004.
28.
Mayer-KuckukP., DoubrovinM., BidautL., Budak-AlpdoganT., CaiS., HubbardV., AlpdoganO., van den BrinkM., BertinoJ.R., BlasbergR.G., BanerjeeD., GelovaniJ.Molecular imaging reveals skeletal engraftment sites of transplanted bone marrow cells. Cell Transplant, 15:75. 2006.
RundleC.H., WangH., YuH., ChadwickR.B., DavisE.I., WergedalJ.E., LauK.H., MohanS., RyabyJ.T., BaylinkD.J.Microarray analysis of gene expression during the inflammation and endochondral bone formation stages of rat femur fracture repair. Bone, 38:521. 2006.
38.
NakazawaT., NakajimaA., SekiN., OkawaA., KatoM., MoriyaH., AmizukaN., EinhornT.A., YamazakiM.Gene expression of periostin in the early stage of fracture healing detected by cDNA microarray analysis. J Orthop Res, 22:520. 2004.
39.
ShimizuT., MehdiR., YoshimuraY., YoshikawaH., NomuraS., MiyazonoK., TakaokaK.Sequential expression of bone morphogenetic protein, tumor necrosis factor, and their receptors in bone-forming reaction after mouse femoral marrow ablation. Bone, 23:127. 1998.
40.
EinhornT.A.The cell and molecular biology of fracture healing. Clin Orthop Relat Res, 355,Suppl:S7. 1999.
41.
JingushiS., JoyceM.E., BolanderM.E.Genetic expression of extracellular matrix proteins correlates with histologic changes during fracture repair. J Bone Miner Res, 7:1045. 1992.
42.
WozneyJ.M., RosenV., CelesteA.J., MitsockL.M., WhittersM.J., KrizR.W., HewickR.M., WangE.A.Novel regulators of bone formation: molecular clones and activities. Science, 242:1528. 1988.
43.
WangE.A., RosenV., D'AlessandroJ.S., BauduyM., CordesP., HaradaT., IsrealD.I., HewickR.M., KernsK.M., LaPanP.et al.Recombinant human bone morphogenetic protein induces bone formation. Proc Natl Acad Sci USA, 87:2220. 1990.
44.
OkuboY., BesshoK., FujimuraK., KonishiY., KusumotoK., OgawaY., IizukaT.Osteoinduction by recombinant human bone morphogenetic protein-2 at intramuscular, intermuscular, subcutaneous and intrafatty sites. Int J Oral Maxillofac Surg, 29:62. 2000.
45.
ZilbermanY., KallaiI., GafniY., PelledG., KossodoS., YaredW., GazitD.Fluorescence molecular tomography enables in vivo visualization and quantification of nonunion fracture repair induced by genetically engineered mesenchymal stem cells. J Orthop Res, 26:522. 2008.
