Asai D J
Dev Biol. 1986 Dec;118(2):416-24. doi: 10.1016/0012-1606(86)90012-6.
Unfertilized sea urchin eggs contain one or more dynein-like enzymes which may be able to serve as microtubule translocators during embryonic development. There are at least two interesting possibilities for the function of the egg dynein: the enzyme may be involved in cytoplasmic microtubule movement such as mitotic spindle anaphase motion; or the enzyme may be a stored precursor for the dynein that functions in embryonic cilia, which are expressed and highly motile at the blastula stage of development. In order to determine directly the distribution and possible function of one of the previously described egg dyneins, the latent-activity 20 S egg dynein (Asai and Wilson, 1985), an antiserum was produced which was highly reactive with the important high Mr polypeptides of 20 S dynein. This antiserum reacted in "Western" immunoblots and in dot-blotting experiments with egg dynein and with embryonic ciliary dynein, but it did not react with any component of sperm flagella. Indirect double immunofluorescence microscopy demonstrated that the anti-20 S antiserum could brightly stain embryonic cilia but it did not stain the sperm flagella from the same sea urchin species. Under the same conditions that the antiserum stained cilia, anti-20 S did not stain the mitotic apparatus but it did appear to stain the cortical region of the dividing egg. In a time-course experiment, the antigen reactive with the anti-20 S antiserum gradually accumulated in the developing early sea urchin embryo. The most significant increase in the apparent concentration of the 20 S dynein occurred just prior to embryonic ciliation and during a period when the mitotic activity of the embryo was in decline. These results lead to two conclusions. First, ciliary dynein and sperm flagellar dynein, although derived from very similar organelles and from the same species of sea urchin, are immunologically distinct. Second, the 20 S egg dynein may be a stored precursor of embryonic ciliary dynein and does not appear to be a component of the mitotic apparatus.
未受精的海胆卵含有一种或多种动力蛋白样酶,在胚胎发育过程中,这些酶可能充当微管转运蛋白。卵动力蛋白的功能至少有两种有趣的可能性:该酶可能参与细胞质微管运动,如有丝分裂纺锤体后期运动;或者该酶可能是胚胎纤毛中发挥作用的动力蛋白的储存前体,胚胎纤毛在发育的囊胚期表达且具有高度运动性。为了直接确定上述卵动力蛋白之一(潜在活性20S卵动力蛋白,浅井和威尔逊,1985年)的分布和可能的功能,制备了一种抗血清,它与20S动力蛋白的重要高Mr多肽具有高度反应性。这种抗血清在“Western”免疫印迹和斑点印迹实验中与卵动力蛋白和胚胎纤毛动力蛋白发生反应,但不与精子鞭毛的任何成分发生反应。间接双重免疫荧光显微镜检查表明,抗20S抗血清可使胚胎纤毛明亮染色,但不能使同一海胆物种的精子鞭毛染色。在抗血清使纤毛染色的相同条件下,抗20S不能使有丝分裂装置染色,但似乎能使正在分裂的卵皮质区域染色。在一项时间进程实验中,与抗20S抗血清反应的抗原在发育中的早期海胆胚胎中逐渐积累。20S动力蛋白表观浓度的最显著增加发生在胚胎纤毛形成之前以及胚胎有丝分裂活性下降的时期。这些结果得出两个结论。第一,纤毛动力蛋白和精子鞭毛动力蛋白,尽管源自非常相似的细胞器且来自同一海胆物种,但在免疫学上是不同的。第二,20S卵动力蛋白可能是胚胎纤毛动力蛋白的储存前体,似乎不是有丝分裂装置的组成部分。