Kimber Susan J, Bagley Peter R
Experimental Embryology and Teratology Unit, MRC Laboratories, SM5 4EF, Carshalton, Surrey, UK.
Rouxs Arch Dev Biol. 1987 Dec;196(8):492-498. doi: 10.1007/BF00399873.
The destination of H-fucose was examined in preimplantation mouse embryos at the 8- to 16-cell stage by autoradiography of semi-thin sections. At the end of a 16-18 h incubation in medium containing 200 μCi/ml L-[6-H] fucose the density of silver grains over the combined surface of cells where they faced their neighbours was 40% greater than that over the outer polar cell surface and 80% greater than that over the cytoplasm. These differences were statistically significant (p<0.01). The outer cell surface also had a grain density above that found over the cytoplasm (p<0.01). There was a rapid decrease of 18-24% in the incorporated radioactivity during the first 1.5 to 3 h in 'chase' medium containing 10 mM 'cold' fucose. Following this the grain density remained relatively stable until it decreased further between 24 and 48 h. By 48 h there was no significant difference between the density of grains over cytoplasm, apposed cell surfaces and outer cell surface: all were reduced compared to the initial values, to 29% in the case of the surfaces and 10% in the case of the cytoplasm. These results indicate that there is an enrichment in the incorporation of H-fucose into macromolecules at the cell surfaces where neighbouring cells confront one another relative to the outer polar cell surface, as well as the cytoplasm, at the 8- to 16-cell stage. Conclusions about the relative density of fucoconjugates per unit length of apposed and polar membrane cannot be reached from such a light microscopic study. Fucoconjugates appear to belong to two groups: the first are turned over relatively rapidly (within 3 h after removal from radioactive medium) while the second are more stable, decaying between 24 and 48 h after removal from H-fucose.
通过对半薄切片进行放射自显影,研究了8至16细胞期植入前小鼠胚胎中H-岩藻糖的去向。在含有200μCi/ml L-[6-H]岩藻糖的培养基中孵育16 - 18小时后,细胞相互邻接表面上的银粒密度比外极细胞表面高40%,比细胞质高80%。这些差异具有统计学意义(p<0.01)。外细胞表面的银粒密度也高于细胞质(p<0.01)。在含有10mM“冷”岩藻糖的“追踪”培养基中,最初1.5至3小时内,掺入的放射性迅速下降18 - 24%。在此之后,银粒密度保持相对稳定,直到24至48小时之间进一步下降。到48小时时,细胞质、邻接细胞表面和外细胞表面的银粒密度之间没有显著差异:与初始值相比均有所降低,表面降低到29%,细胞质降低到10%。这些结果表明,在8至16细胞期,相对于外极细胞表面和细胞质,在相邻细胞相互面对的细胞表面,H-岩藻糖掺入大分子的量有所增加。从这样的光学显微镜研究中无法得出关于邻接膜和极膜每单位长度岩藻糖结合物相对密度的结论。岩藻糖结合物似乎分为两组:第一组周转相对较快(从放射性培养基中取出后3小时内),而第二组更稳定,在从H-岩藻糖中取出后24至48小时之间降解。