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Cytochemical demonstration of the glucocorticoid receptor in skeletal cells of the rat.

机译:大鼠骨骼细胞中糖皮质激素受体的细胞化学证明。

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The in vivo localization of the glucocorticoid receptor (GR) was studied in cartilage and bone cells of femurs of young adult rats. Deparaffinized sections were treated with a polyclonal antibody raised against the amino-terminus of human GR; the immunoreaction was detected with the streptavidin-biotin amplification method. Histomorphometric, computer-assisted analysis of GR-positive cells was performed by counting the percentage of GR-immunostained cells in the proliferative and maturative/hypertrophic zone of the epiphyseal growth plate cartilage, and of the percentage of positive osteoblasts (OBs), osteoclasts (OCLS) and osteocytes (OCs) in the metaphyseal secondary ossification zone. Numbers of OBs and OCLs per mm of metaphyseal endosteal perimeter, and numbers of OCs per mm2 of trabecular area were also counted. Immunopositive cells were found both in cartilage and bone, with variable degree of nuclear and/or cytoplasmic immunostaining; immunonegative cells were present among the positive ones. Histomorphometry showed that about 54% of chondrocytes in the proliferative zone, and 55% of chondrocytes in the maturative/hypertrophic zone of the growth plate were labeled; in metaphyseal bone, 68% of OBs, 65% of OCs, and 98% of OCLs were GR-positive. The density of positive cells was 12.06 OBs/mm, 3.32 OCLs/mm, and 520.40 OCs/mm2. These results, for the first time obtained in vivo, show that GR is present in cartilage and bone cells, and that the degree of GR-immunostaining is variable in the same type of cell. This may be dependent on the cell cycle and stage of differentiation, and may reflect a variable cellular sensitivity to the stimulation of the glucocorticoid hormone.
机译:研究了成年大鼠股骨软骨和骨细胞中糖皮质激素受体(GR)的体内定位。用针对人GR氨基末端的多克隆抗体处理去石蜡切片。用链霉亲和素-生物素扩增法检测免疫反应。通过计数骨epi生长板软骨增生和成熟/肥大区的GR免疫染色细胞的百分比以及阳性成骨细胞(OBs),破骨细胞( OCLS)和干phy端次生骨化区的骨细胞(OCs)。还计算了每毫米of骨干内膜骨膜周长的OB和OCL数,以及每mm2小梁面积的OCs数。在软骨和骨骼中均发现了免疫阳性细胞,具有不同程度的核和/或细胞质免疫染色。阳性细胞中存在免疫阴性细胞。组织形态计量学显示,在生长板的增生区中约有54%的软骨细胞被标记,在生长板的成熟/肥大区中有约55%的软骨细胞被标记。在干phy端骨中,68%的OB,65%的OC和98%的OCL为GR阳性。阳性细胞的密度为12.06 OBs / mm,3.32 OCLs / mm和520.40 OCs / mm2。这些结果首次在体内获得,表明GR存在于软骨和骨细胞中,并且GR免疫染色的程度在同一类型的细胞中是可变的。这可能取决于细胞周期和分化阶段,并且可能反映出对糖皮质激素刺激的可变细胞敏感性。

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