Petit B, Ronzière M C, Hartmann D J, Herbage D
Institut de Biologie et Chimie des Proteines (I.B.C.P.), UPR C.N.R.S. 412, Lyon, France.
Histochemistry. 1993 Sep;100(3):231-9. doi: 10.1007/BF00269096.
Type XI collagen was localized with polyclonal antibodies specific for alpha 1 (XI) and alpha 2 (XI) chains in the resting zone of epiphyseal cartilage from calf fetuses. The immunofluorescence technique was used on sections of cartilage, and the immunogold labelling technique for electron microscopy on fibrils isolated from cartilage and, for the first time, in situ on blocks of cartilage fractured in liquid nitrogen. Immunofluorescence showed that without pepsin treatment the staining of type XI collagen was restricted to the pericellular zones; after pepsin treatment, the staining was co-distributed with that of type II collagen. Immunoelectron microscopy performed on isolated fibrils and on cartilage blocks showed that after disruption of fibrils with pepsin, type XI collagen was labelled on small filaments on the fibrils. When the fibrils were not disrupted, labelling was observed in situ only at the ends of the fibrils or on cross-sections of some fibrils. These results indicate that type XI collagen is located inside type II collagen fibrils in fetal bovine epiphyseal cartilage, as already postulated for embryonic chicken sterna.
利用针对α1(XI)和α2(XI)链的多克隆抗体,在胎牛骺软骨的静止区定位了XI型胶原蛋白。对软骨切片采用免疫荧光技术,对从软骨中分离出的原纤维以及首次在液氮中破碎的软骨块原位采用免疫金标记技术进行电子显微镜观察。免疫荧光显示,未经胃蛋白酶处理时,XI型胶原蛋白的染色仅限于细胞周围区域;经胃蛋白酶处理后,染色与II型胶原蛋白的染色共同分布。对分离出的原纤维和软骨块进行的免疫电子显微镜观察表明,用胃蛋白酶破坏原纤维后,XI型胶原蛋白在原纤维上的小细丝上被标记。当原纤维未被破坏时,仅在原纤维的末端或一些原纤维的横截面上观察到原位标记。这些结果表明,XI型胶原蛋白位于胎牛骺软骨的II型胶原原纤维内部,这与胚胎鸡胸骨的情况一致。