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 bi...

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Bibliographic Details
Published in:Journal of molecular histology 2000-03, Vol.32 (3), p.175-186
Main Authors: Takagi, M, Kamiya, N, Urushizaki, T, Tada, Y, Tanaka, H
Format: Article
Language:English
Subjects:
Animals
Animals, Newborn
Biglycan
Blotting, Northern
Calcification, Physiologic
Extracellular Matrix - ultrastructure
Extracellular Matrix Proteins
Gene Expression
Growth Plate - ultrastructure
Immunohistochemistry
In Situ Hybridization
Microscopy, Immunoelectron
Proteins
Proteoglycans - genetics
Proteoglycans - isolation & purification
Rats
Rats, Wistar
RNA, Messenger - isolation & purification
Rodents
Space life sciences
Tibia - ultrastructure
Tissue Distribution
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Summary: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.
ISSN:0018-2214
1567-2379
1567-2387
1573-6865
DOI:10.1023/A:1004099324169