Bioluminescence imaging of bone formation using hairless osteocalcin-luciferase transgenic mice.
Osteocalcin is a major noncollagenous protein component of bone extracellular matrix, synthesized and secreted exclusively by osteoblastic cells during the late stage of maturation. We introduced a 10kb human osteocalcin enhancer/promoter (OC)-luciferase (Luc) construct into a hairless mouse line. Examination of tissue RNAs from these transgenic mice showed a predominant restriction of Luc mRNA expression to bone-associated tissues. Immunohistochemical staining of calvaria tissue sections revealed the localization of Luc protein to osteoblasts. Utilizing in vivo bioluminescence imaging, supplementation of 1α,25-dihydroxyvitamin D(3) increased Luc activity throughout the skeleton, consistent with in vitro transient transfection studies in osteoblast-like cells. Moreover, we observed an abrupt decrease in bioluminescence activity as the mice reached puberty, and a further decrease gradually thereafter. Using a radius skeletal repair model, we observed enhanced bioluminescence at the fracture site in both young (14-22 weeks old) and aged (50-66 weeks old) mice. However, peak bioluminescence was delayed in aged mice compared with young mice, suggesting retarded osteocalcin expression with aging. Our in vivo imaging system may contribute to the therapy and prevention of various bone metabolic disorders through its effective monitoring of the bone formation process.
Division of Molecular Biology, School of Life Sciences, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8503, Japan. tomo@med.tottori-u.ac.jp
Tomoko
T
Nakanishi
Kazuo
K
Kokubun
Haruka
H
Oda
Mika
M
Aoki
Atsumi
A
Soma
Makoto
M
Taniguchi
Yasuhiro
Y
Kazuki
Mitsuo
M
Oshimura
Kenzo
K
Sato
D000375:Aging / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D017102:Fracture Healing; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008156:Luciferases / Q000378:metabolism*; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D008812:Mice, Hairless; D008822:Mice, Transgenic; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism*; D010012:Osteogenesis; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics
D015675:Osteocalcin; D008156:Luciferases