Jingushi S, Scully S P, Joyce M E, Sugioka Y, Bolander M E
Department of Orthopaedic Surgery, Kyushu University, Fukuoka, Japan.
J Orthop Res. 1995 Sep;13(5):761-8. doi: 10.1002/jor.1100130516.
Chondrocytes in the growth plate progress in an orderly fashion from resting through proliferating to hypertrophic cells. In the region of hypertrophic chondrocytes, the cartilage is invaded by capillary loops and endochondral ossification is initiated. It is currently believed that growth factors may regulate the proliferation and maturation of chondrocytes and the synthesis of extracellular matrix in the growth plate. The ordered sequence of proliferation and differentiation observed in the growth plate provides a unique opportunity to study the role of acidic fibroblast growth factor, basic fibroblast growth factor, and transforming growth factor-beta 1 in the regulation of these processes. In this study, expression of the mRNA of these growth factors was examined using total RNA extracted from the physis and epiphysis of rat tibias. Transforming growth factor-beta 1 mRNA was detected by Northern hybridization. Expression of the genes encoding acidic and basic fibroblast growth factors was demonstrated by polymerase chain reaction amplification. In addition, using polyclonal antibodies against these growth factors, we localized them by immunohistochemical analysis. Strong intracellular staining with a predominantly nuclear pattern was observed in chondrocytes from the proliferating and upper hypertrophic zones. In contrast, chondrocytes in the resting zone stained only faintly for the presence of these growth factors. Some chondrocytes in the resting zone adjacent to the proliferating zone stained with these antibodies, and the antibodies also stained cells in the zone of Ranvier, which regulates latitudinal bone growth. Lastly, the location of transforming growth factor-beta 1 was examined further with use of a polyclonal antipeptide antibody specific for its extracellular epitope.(ABSTRACT TRUNCATED AT 250 WORDS)
生长板中的软骨细胞以有序的方式从静止状态发展为增殖状态,再到肥大细胞状态。在肥大软骨细胞区域,软骨被毛细血管袢侵入,软骨内成骨开始。目前认为,生长因子可能调节软骨细胞的增殖和成熟以及生长板中细胞外基质的合成。生长板中观察到的增殖和分化的有序序列为研究酸性成纤维细胞生长因子、碱性成纤维细胞生长因子和转化生长因子-β1在这些过程调节中的作用提供了独特的机会。在本研究中,使用从大鼠胫骨的干骺端和骨骺提取的总RNA检测了这些生长因子的mRNA表达。通过Northern杂交检测转化生长因子-β1 mRNA。通过聚合酶链反应扩增证实了编码酸性和碱性成纤维细胞生长因子的基因的表达。此外,使用针对这些生长因子的多克隆抗体,通过免疫组织化学分析对它们进行定位。在增殖区和上肥大区的软骨细胞中观察到强烈的细胞内染色,主要呈核模式。相比之下,静止区的软骨细胞对这些生长因子的染色仅很淡。与增殖区相邻的静止区的一些软骨细胞用这些抗体染色,这些抗体也对调节横向骨生长的朗飞氏区的细胞进行染色。最后,使用对其细胞外表位具有特异性的多克隆抗肽抗体进一步研究了转化生长因子-β1的定位。(摘要截短至250字)