Chadwick C M
Department of Biological Sciences University of Pittsburgh Pittsburgh, PA 15260 USA.
Dev Growth Differ. 1986 Apr;28(2):203-211. doi: 10.1111/j.1440-169X.1986.00203.x.
The developmental regulation of the vegetative cohesion molecule, gp 126, has been monitored in the cellular slime mould Dictyostelium discoideum. As judged by immunoprecipitation using an anti-vegetative cell, cohesion blocking antibody, gp126 persisted until at least the grex stage of development although a decline in the level of the molecule was observed thereafter. Further, after the grex stage, cells showed an increasing loss of ability to absorb the cohesion-blocking effect of an anti-vegetative cell Fab. Therefore, the decline of gp 126 could be ascribed to a loss from the cell surface. By radio-iodination of vegetative cells followed by liquid-scintillation counting of gp 126, developmental-regulation could be determined quantitatively. At the grex stage of development, whole aggregates were embedded in wax. Longitudinal sections were then stained with a monospecific anti-gp 126 Fab, followed by a fluorescent sheep anti-rabbit IgG. Fluorescence was observed only at the tip (the prestalk) region, thereby showing that gp 126 is a prestalk marker. To confirm the above result, grexes were dissociated and cells were separated into prespore and prestalk populations on a gradient of Percoll. Prestalk but not prespore cells were able to absorb the cohesion-blocking effect of an anti-vegetative cell Fab. To examine the biosynthesis of gp 126, cells were pulsed with radioactive glucosamine, mannose or acetate. The pattern of incorporation of radioactivity suggested that the de novo synthesis of gp 126 ceases upon commencement of development.
在细胞黏菌盘基网柄菌中,已对营养性黏附分子gp 126的发育调控进行了监测。使用抗营养细胞的黏附阻断抗体进行免疫沉淀判断,gp126至少持续到发育的蛞蝓体阶段,不过此后观察到该分子水平有所下降。此外,在蛞蝓体阶段之后,细胞对吸收抗营养细胞Fab黏附阻断作用的能力逐渐丧失。因此,gp 126的下降可归因于从细胞表面的丢失。通过对营养细胞进行放射性碘化,然后对gp 126进行液体闪烁计数,可以定量确定发育调控情况。在发育的蛞蝓体阶段,将整个聚集体包埋在蜡中。然后将纵切片用单特异性抗gp 126 Fab染色,接着用荧光羊抗兔IgG染色。仅在顶端(前柄)区域观察到荧光,从而表明gp 126是一种前柄标记物。为了证实上述结果,将蛞蝓体解离,并在Percoll梯度上把细胞分离成前孢子和前柄群体。前柄细胞而非前孢子细胞能够吸收抗营养细胞Fab的黏附阻断作用。为了研究gp 126的生物合成,用放射性葡糖胺、甘露糖或乙酸盐对细胞进行脉冲标记。放射性掺入模式表明,gp 126的从头合成在发育开始时停止。