Hayashi M, Hayashi K, Iyama K, Trelstad R L, Linsenmayer T F, Mayne R
Department of Pathology, UMDNJ-Robert Wood Johnson Medical School, Piscataway 08854.
Dev Dyn. 1992 Jul;194(3):169-76. doi: 10.1002/aja.1001940302.
The notochord of embryonic chicks produces type IX collagen, as well as type II collagen, prior to the onset of vertebral chondrogenesis. To address the question of whether the notochord secretes the "long-form" type IX collagen found in cartilage or the "short-form" type IX found in the cornea and vitreous humor, we examined immunoreactivity of the notochordal type IX collagen using two different monoclonal antibodies. The antibody 2C2 recognizes an epitope close to the carboxyl-terminus of the HMW fragment, which is present in both the long- and short-form type IX collagens, whereas another antibody 4D6 recognizes an epitope in the NC4 domain of the long-form type IX collagen, which is absent in the short-form type IX collagen. Therefore, the long-form is recognized by its reaction with both 2C2 and 4D6, while the short-form by its reaction with only 2C2 and no reaction with 4D6. Immunostaining of vertebral sections with 2C2 shows an identical distribution of staining with that for type II collagen, although the staining with 2C2 is less intense. The 2C2-reactive type IX collagen is found within the notochord at stage 14 and in the notochordal sheath at stage 20. Deposition of this collagen in the perinotochordal matrix increases with time and reaches a level comparable with that for type II at stage 31. In contrast, the 4D6-reactive type IX collagen is not found within the notochord nor in the notochordal sheath.(ABSTRACT TRUNCATED AT 250 WORDS)
在脊椎软骨形成开始之前,胚胎小鸡的脊索会产生IX型胶原蛋白以及II型胶原蛋白。为了探究脊索分泌的是软骨中发现的“长形式”IX型胶原蛋白,还是角膜和玻璃体液中发现的“短形式”IX型胶原蛋白,我们使用两种不同的单克隆抗体检测了脊索IX型胶原蛋白的免疫反应性。抗体2C2识别靠近HMW片段羧基末端的一个表位,该表位存在于长形式和短形式的IX型胶原蛋白中,而另一种抗体4D6识别长形式IX型胶原蛋白NC4结构域中的一个表位,该表位在短形式IX型胶原蛋白中不存在。因此,长形式通过与2C2和4D6的反应被识别,而短形式仅通过与2C2的反应被识别,与4D6无反应。用2C2对椎骨切片进行免疫染色显示,其染色分布与II型胶原蛋白相同,尽管2C2的染色强度较低。在第14阶段,2C2反应性IX型胶原蛋白存在于脊索内,在第20阶段存在于脊索鞘中。这种胶原蛋白在脊索周围基质中的沉积随时间增加,并在第31阶段达到与II型胶原蛋白相当的水平。相比之下,在脊索内和脊索鞘中均未发现4D6反应性IX型胶原蛋白。(摘要截短至250字)