Callebaut Marc, Van Nueten Emmy, Bortier Hilde, Harrisson Fernand
Laboratory of Human Anatomy and Embryology UA RUCA, B-2020 Antwerpen, Belgium.
J Morphol. 2002 Aug;253(2):132-47. doi: 10.1002/jmor.1116.
Using the quail-chick chimera technique, we followed the fate of Rauber's sickle cells in older whole blastoderms (cultured for approximately 2 days): after removal of the autochthonous Rauber's sickle from an unincubated chicken blastoderm, a quail Rauber's sickle was grafted isotopically and isochronically in its place. In transverse sections through these chimeras, the grafted quail Rauber's sickle cells were seen to have transformed into a broad row or ridge of quail junctional endoblast cells extending at the inner border of the area containing blood islands. After unilateral removal of the junctional endoblast from an intermediate streak chicken blastoderm (Stage 3; Hamburger and Hamilton [1951] J Morphol 88:49-92), we observed during further in vitro culture that at the operated side, in the area previously occupied by this junctional endoblast, blood islands no longer developed. If after such a unilateral removal of the chicken junctional endoblast quail junctional endoblast was apposed in its place, then blood islands reappeared in the operated area. The intimate contact between the apposed quail junctional endoblast and the recently formed blood islands, derived from peripherally migrating mesoderm, was very obvious on sections through such chimeras. We further demonstrate that Rauber's sickle vs. junctional endoblast is indispensable for the anlage of blood islands in avian blastoderms. Indeed, in the absence of Rauber's sickle material no blood islands develop (even when mesoderm is present after ingression of the upper layer via a primitive streak) in the isolated central region of the area centralis of unincubated chicken blastoderms after culture in vitro. Also, no junctional endoblast and no sickle canal appear in these explants. By contrast, if a Rauber's sickle fragment is placed on such an isolated central blastoderm region, then blood islands develop. These blood islands start to develop from peripherally migrating mesoderm in the neighborhood of the Rauber's sickle-derived junctional endoblast.
利用鹌鹑 - 鸡嵌合体技术,我们追踪了较老的完整胚盘(培养约2天)中劳伯氏镰刀细胞的命运:从未孵化的鸡胚盘中移除自体的劳伯氏镰刀后,将鹌鹑的劳伯氏镰刀等部位、等时间地移植到其位置。在这些嵌合体的横切面上,可以看到移植的鹌鹑劳伯氏镰刀细胞已转化为一排宽阔的鹌鹑连接内胚层细胞或嵴,在含有血岛的区域内边界处延伸。在从中间条带鸡胚盘(第3阶段;汉堡和汉密尔顿[1951]《形态学杂志》88:49 - 92)单侧移除连接内胚层后,我们在进一步的体外培养过程中观察到,在手术侧,在先前被该连接内胚层占据的区域,血岛不再发育。如果在单侧移除鸡连接内胚层后将鹌鹑连接内胚层置于其位置,那么血岛会在手术区域重新出现。在通过此类嵌合体的切片上,相邻的鹌鹑连接内胚层与源自周边迁移中胚层的新形成血岛之间的紧密接触非常明显。我们进一步证明,劳伯氏镰刀与连接内胚层对于鸟类胚盘血岛的原基形成是不可或缺的。实际上,在未孵化的鸡胚盘中央区的分离中央区域,体外培养后,若没有劳伯氏镰刀物质,即使在上层通过原条内迁后存在中胚层,也不会形成血岛。此外,这些外植体中也不会出现连接内胚层和镰刀管。相比之下,如果将一块劳伯氏镰刀碎片放置在这样一个分离的中央胚盘区域,那么血岛就会发育。这些血岛开始从劳伯氏镰刀衍生的连接内胚层附近的周边迁移中胚层发育而来。