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Quantitative detection of anionic sites in rat femoral cartilage using
cationic colloidal gold at low pH levels
H. Ueda, Y. Kato and S. Ohno
Department of Anatomy, Yamanashi Medical University, Tamaho,
Yamanashi, Japan
Offprint requests to: Dr. Shinichi
Ohno, MD, PhD., Professor and Chairman, Department of Anatomy, Yamanashi
Medical University, 1110 Shimokato, Tamaho, Yamanashi 409-3898, Japan. Fax:
81-552-73-6743
Summary. Quantitative analyses
of anionic sites in the rat cartilage were performed by using post-embedding
LR-Gold method with cationic colloidal gold (CCG) at different pH levels.
Alteration of the pH levels affected the CCG staining patterns. The CCG
particles were more observed at low pH levels, 1.0 and 2.5, than those at
high pH 7.4. So, we quantitatively examined the superficial, intermediate
and deep layers at pH 1.0. Morphometric analyses showed that CCG particles
were increased from the superficial layer to the deep layer in both pericellular
matrix (PM) and interterritorial matrix (IM). Moreover, they were more observed
in PM than in IM. After chondroitinase ABC digestion, the CCG labeling was
reduced in all matrices of the superficial and intermediate layers, though
many CCG particles were still observed in PM of the deep layer. After hyaluronidase
digestion, the CCG labeling was markedly reduced in the PM of the deep layer.
These findings suggest that anionic sites in the cartilage matrix are detected
by using the CCG only at low pH levels. In addition, the PM in the deep
layer may consist of different components from those of the other two layers.
Histol. Histopathol. 13, 1001-1009 (1998)
Key words: Hyaline cartilage,
Catinonic colloidal gold, pH level, Proteoglycan, Anionic site
DOI: 10.14670/HH-13.1001
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