Gene expression and immunohistochemical localization of biglycan in association with mineralization in the matrix of epiphyseal cartilage.
This study has used in situ hybridization, Northern blot analysis, and immunohistochemistry at the light and electron microscope levels to localize mRNAs and core proteins of biglycan in developing tibial epiphyseal cartilage of 10-day old Wistar rats. The expression of mRNAs and core proteins of biglycan appeared prominent in hypertrophic and degenerative chondrocytes associated with the epiphyseal ossification centre and the growth plate cartilage, but was not seen in the rest of epiphyseal cartilage. Northern blot analysis confirmed biglycan mRNA expression in the epiphyseal cartilage. Ultrastructural immunogold cytochemistry of the growth plate revealed that prominent immunolabelling was confined to the Golgi apparatus and cisternae of rough-surfaced endoplasmic reticulum of the hypertrophic and the degenerating chondrocytes, the early mineralized cartilage matrices of the longitudinal septum of the lower hypertrophic and the calcifying zones, and fully mineralized cartilage mitrices, which were present in the metaphyseal bone trabeculae. Furthermore, Western blot analysis of biglycan in extracts of fresh epiphyseal cartilage revealed that an EDTA extract, after chondroitinase ABC digestion, contains core proteins of biglycan, indicating the presence of biglycan in mineralized cartilage matrices. These results indicate that the distribution of biglycan is associated with cartilage matrix mineralization.
Takagi
M
M
Department of Anatomy, Nihon University School of Dentistry, Tokyo, Japan.
Kamiya
N
N
Urushizaki
T
T
Tada
Y
Y
Tanaka
H
H
D000818:Animals; D000831:Animals, Newborn; D058578:Biglycan; D015152:Blotting, Northern; D002113:Calcification, Physiologic; D005109:Extracellular Matrix / Q000648:ultrastructure*; D016326:Extracellular Matrix Proteins; D015870:Gene Expression; D006132:Growth Plate / Q000648:ultrastructure*; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D016253:Microscopy, Immunoelectron; D011509:Proteoglycans / Q000235:genetics / Q000302:isolation & purification*; D012333:RNA, Messenger / Q000302:isolation & purification; D051381:Rats; D017208:Rats, Wistar; D013977:Tibia / Q000648:ultrastructure; D014018:Tissue Distribution
D058578:Biglycan; D016326:Extracellular Matrix Proteins; D011509:Proteoglycans; D012333:RNA, Messenger