A continuous observation of the degenerative process in the intervertebral disc of Smad3 gene knock-out mice.
METHODS
Pathologic changes were observed in the spine of small mother against decapentaplegic (Smad) 3 mice at different time points.


OBJECTIVE
To observe the degeneration of the intervertebral disc (IVD) in Smad3 gene knock-out mice with growth.


BACKGROUND
Smad3 gene knock-out (Smad3) mice displays phenotypes similar to human osteoarthritis. Despite the similarities between IVD cartilage endplate and the articular cartilage, there has been relatively little interest in exploring the possibility that IVD degeneration might be driven by the deficiency of Smad3.


METHODS
The Smad3 mice were killed at the 10th, 30th, and 60th day after their birth and the IVD samples of spine were harvested for histologic and immunohistochemical studies. Total RNA isolated from these samples were used for real-time PCR analysis of type II collagen (Col2alpha1), type X collagen (Col10alpha1), aggrecan, and transforming growth factor-beta1 (TGF-beta1).


RESULTS
Compared with the wild-type mice, Smad3 mice appeared significantly smaller in size. Radiograph showed that the spine of Smad3 mice is malformation and kyphosis. Histologic analysis revealed the declined height of cartilage endplate, decreased proteoglycan and collagen content in disc of Smad3 mice. With growth, especially of the 30- and 60-day old Smad3 mice, the protein positive staining of type II collagen, aggrecan, and TGF-beta1 in the disc decreased, while that of type X collagen increased. And the analysis of real-time PCR showed that the mRNA expression of Col2alpha1, aggrecan, and TGF-beta1 decreased, while that of Col10alpha1 increased.


CONCLUSIONS
Smad3 gene knock-out mice develop IVD degeneration with growth.
Li
Chen-Guang
CG
Institute of Spine, Shanghai University of Traditional Chinese Medicine, Shanghai, People's Republic of China.
Liang
Qian-Qian
QQ
Zhou
Quan
Q
Menga
Emmanuel
E
Cui
Xue-Jun
XJ
Shu
Bing
B
Zhou
Chong-Jian
CJ
Shi
Qi
Q
Wang
Yong-Jun
YJ
D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D000818:Animals; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D024121:Collagen Type X / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007403:Intervertebral Disc / Q000378:metabolism / Q000473:pathology*; D007405:Intervertebral Disc Displacement / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D010003:Osteoarthritis / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051900:Smad3 Protein / Q000235:genetics / Q000502:physiology*; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism
D053674:Aggrecans; C447413:Col2a1 protein, mouse; D024043:Collagen Type II; D024121:Collagen Type X; D012333:RNA, Messenger; D051900:Smad3 Protein; C497791:Smad3 protein, mouse; D053773:Transforming Growth Factor beta1