Levi G, Crossin K L, Edelman G M
Rockefeller University, New York 10021.
J Cell Biol. 1987 Nov;105(5):2359-72. doi: 10.1083/jcb.105.5.2359.
Studies of chicken embryos have demonstrated that cell adhesion molecules are important in embryonic induction and are expressed in defined sequences during embryogenesis and histogenesis. To extend these observations and to provide comparable evidence for heterochronic changes in such sequences during evolution, the local distributions of the neural cell adhesion molecule (N-CAM) and of the liver cell adhesion molecule (L-CAM) were examined in Xenopus laevis embryos by immunohistochemical and biochemical techniques. Because of the technical difficulties presented by the existence of multiple polypeptide forms of CAMs and by autofluorescence of yolk-containing cells, special care was taken in choosing and characterizing antibodies, fluorophores, and embedding procedures. Both N-CAM and L-CAM were found at low levels in pregastrulation embryos. During gastrulation, N-CAM levels increased in the presumptive neural epithelium and decreased in the endoderm, but L-CAM continued to be expressed in all cells including endodermal cells. During neurulation, the level of N-CAM expression in the neural ectoderm increased considerably, while remaining constant in non-neural ectoderm and diminishing in the somites; in the notochord, N-CAM was expressed transiently. Prevalence modulation was also seen at all sites of secondary induction: both CAMs increased in the sensory layer of the ectoderm during condensation of the placodes. During organogenesis, the expression of L-CAM gradually diminished in the nervous system while N-CAM expression remained high. In all other organs examined, the amount of one or the other CAM decreased, so that by stage 50 these two molecules were expressed in non-overlapping territories. Embryonic and adult tissues were compared to search for concordance of CAM expression at later stages. With few exceptions, the tissue distributions of N-CAM and L-CAM were similar in the frog and in the chicken from early times of development. In contrast to previous observations in the chicken and in the mouse, N-CAM expression was found to be high in the adult liver of Xenopus, whereas L-CAM expression was low. In the adult brain, N-CAM was expressed as three components of apparent molecular mass 180, 140, and 120 kD, respectively; in earlier stages of development only the 140-kD component could be detected. In the liver, a single N-CAM band appears at 160 kD, raising the possibility that this band represents an unusual N-CAM polypeptide. L-CAM appeared at all stages as a 124-kD molecule.(ABSTRACT TRUNCATED AT 400 WORDS)
对鸡胚胎的研究表明,细胞黏附分子在胚胎诱导中起重要作用,并且在胚胎发生和组织发生过程中按特定顺序表达。为了扩展这些观察结果,并为进化过程中此类序列的异时性变化提供可比证据,通过免疫组织化学和生化技术,研究了非洲爪蟾胚胎中神经细胞黏附分子(N-CAM)和肝细胞黏附分子(L-CAM)的局部分布。由于细胞黏附分子存在多种多肽形式以及含卵黄细胞的自发荧光带来的技术难题,在选择和鉴定抗体、荧光团及包埋程序时格外小心。在原肠胚形成前的胚胎中,N-CAM和L-CAM的水平都很低。在原肠胚形成期间,N-CAM水平在预定神经上皮中升高,在内胚层中降低,但L-CAM在包括内胚层细胞在内的所有细胞中持续表达。在神经胚形成期间,神经外胚层中N-CAM的表达水平大幅增加,而非神经外胚层中保持不变,在体节中减少;在脊索中,N-CAM短暂表达。在二次诱导的所有部位也观察到了表达水平的调节:在基板凝聚过程中,两种细胞黏附分子在外胚层的感觉层中均增加。在器官发生过程中,L-CAM在神经系统中的表达逐渐减少,而N-CAM的表达仍保持高水平。在所有其他检查的器官中,一种或另一种细胞黏附分子的量减少,因此到第50阶段时,这两种分子在不重叠的区域表达。比较胚胎组织和成年组织以寻找后期细胞黏附分子表达的一致性。除少数例外,从发育早期开始,非洲爪蟾和鸡中N-CAM和L-CAM的组织分布相似。与之前在鸡和小鼠中的观察结果相反,发现非洲爪蟾成年肝脏中N-CAM表达高,而L-CAM表达低。在成年大脑中,N-CAM分别以表观分子量为180、140和120 kD的三种成分表达;在发育早期只能检测到140-kD的成分。在肝脏中,一条单一的N-CAM条带出现在160 kD处,这增加了这条带代表一种不寻常的N-CAM多肽的可能性。L-CAM在所有阶段均表现为124-kD的分子。(摘要截选至400字)