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生长板软骨基质中凝集素结合糖缀合物的原位定位

In situ localization of lectin-binding glycoconjugates in the matrix of growth-plate cartilage.

作者信息

Farnum C E, Wilsman N J

出版信息

Am J Anat. 1986 May;176(1):65-82. doi: 10.1002/aja.1001760106.

Abstract

In the distal hypertrophic zone of growth-plate cartilage, the pericellular matrix surrounding individual chondrocytes and the territorial matrix uniting chondrocytes into columnar groups are invaded by metaphyseal endothelial cells prior to osteogenesis. In the present study, lectin-binding glycoconjugates were analyzed in these two matrix compartments of growth-plate cartilage from Yucatan swine. Nine lectin-fluorescein conjugates were tested by a postembedment method on 1-micron-thick, nondecalcified, Epon-embedded sections. Chondrocytes in all cellular zones were surrounded by a pericellular matrix which showed positive binding for peanut agglutinin (PNA), ricin agglutinin (RCA-I), and soybean agglutinin (SBA). Binding by these lectins was sensitive to digestion with hyaluronidase, chondroitinase, and trypsin. Pericellular glyconconjugtes that bind RCA-I and concanvalin A (CONA) after periodic acid oxidation, and which were sensitive to trypsin but not to chondroitinase or hyaluronidase, were present in the hypertrophic cell zone. Within the territorial matrix, binding of lectins specific for galactose, N-acetylgalactosamine, and fucose showed gradients of intensity which became maximal at the last transverse septum. Lectin-binding histochemistry more precisely differentiated the microheterogeneity of glycoconjugate distribution within these two matrix compartments than has been possible with other histochemical techniques. Lectin-binding affinity is a potentially useful technique by which to isolate cartilage matrix macromolecules unique to specific cellular zones of the growth plate.

摘要

在生长板软骨的远端肥厚区,在成骨之前,干骺端内皮细胞侵入围绕单个软骨细胞的细胞周基质以及将软骨细胞连接成柱状群的区域基质。在本研究中,对尤卡坦猪生长板软骨的这两个基质区室中的凝集素结合糖缀合物进行了分析。通过包埋后方法在1微米厚的、未脱钙的、环氧树脂包埋的切片上测试了九种凝集素 - 荧光素缀合物。所有细胞区的软骨细胞都被细胞周基质包围,该基质对花生凝集素(PNA)、蓖麻凝集素(RCA-I)和大豆凝集素(SBA)显示出阳性结合。这些凝集素的结合对透明质酸酶、软骨素酶和胰蛋白酶的消化敏感。在肥大细胞区存在经高碘酸氧化后结合RCA-I和伴刀豆球蛋白A(CONA)且对胰蛋白酶敏感但对软骨素酶或透明质酸酶不敏感的细胞周糖缀合物。在区域基质内,对半乳糖、N-乙酰半乳糖胺和岩藻糖具有特异性的凝集素结合显示出强度梯度,在最后一个横向隔膜处达到最大值。凝集素结合组织化学比其他组织化学技术更精确地区分了这两个基质区室内糖缀合物分布的微观异质性。凝集素结合亲和力是一种潜在有用的技术,可用于分离生长板特定细胞区独特的软骨基质大分子。

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