Harrisson F, Van Hoof J, Vanroelen C, Vakaet L
Cell Tissue Res. 1985;239(3):643-9. doi: 10.1007/BF00219243.
A chemical basis for the transmission of signals during gastrulation has been investigated by using chimaeric embryos resulting from the combination of 3H-glucosamine-labelled and unlabelled hypoblast with epiblast taken from chicken and quail embryos at stage 3 of Vakaet (1970). The ability to distinguish chicken from quail cells on the basis of their different nuclear distribution of heterochromatin after Feulgen staining made it possible to determine the origin of the cells in the chimaerae. Tritiated quail hypoblast (after incubation of the embryo in the presence of 3H-glucosamine) was transplanted onto unlabelled chicken blastoderm deprived of its hypoblast. After culture of the chimaera for 5 h, the autoradiographic pattern shows silver grains not only over the graft, but also at the ventral surface of the epiblast of the host. Transfer of label may occur to mesoblast cells, but not between chicken and quail hypoblast cells. Chase experiments exclude the possibility that unprocessed, tritiated glucosamine is transferred. Chemical fixation of the host before transplantation of a labelled quail hypoblast also allows visualization of a transfer of macromolecules from hypoblast to the basement membrane of the epiblast, suggesting that an intervention of the epiblast cells in this process is not necessary. The morphology of the chimaeric embryos, as studied by scanning electron microscopy, suggests a direct deposition of these macromolecules by filopodia of the dorsal surface of the hypoblast. The possibility of diffusion of free macromolecules has been considered and can reasonably be discarded on the basis of several observations.(ABSTRACT TRUNCATED AT 250 WORDS)
通过使用嵌合体胚胎,研究了原肠胚形成过程中信号传递的化学基础。这些嵌合体胚胎由3H-葡萄糖胺标记和未标记的下胚层与取自瓦凯特(1970年)第3阶段鸡和鹌鹑胚胎的上胚层组合而成。基于福尔根染色后异染色质在细胞核中的不同分布,能够区分鸡和鹌鹑细胞,这使得确定嵌合体中细胞的来源成为可能。将经氚标记的鹌鹑下胚层(在胚胎于3H-葡萄糖胺存在下孵育后)移植到去除下胚层的未标记鸡胚盘上。将嵌合体培养5小时后,放射自显影片显示银粒不仅出现在移植物上,也出现在宿主上胚层的腹面。标记可能转移到中胚层细胞,但不会在鸡和鹌鹑的下胚层细胞之间转移。追踪实验排除了未加工的氚标记葡萄糖胺转移的可能性。在移植标记的鹌鹑下胚层之前对宿主进行化学固定,也能观察到大分子从下胚层转移到上胚层的基底膜,这表明上胚层细胞在此过程中并非必需。通过扫描电子显微镜研究嵌合体胚胎的形态,表明这些大分子是由下胚层背面的丝状伪足直接沉积的。已经考虑了游离大分子扩散的可能性,基于多项观察结果可以合理地排除这种可能性。(摘要截短于250字)