Callebaut Marc, Van Nueten Emmy, Bortier Hilde, Harrisson Fernand
Laboratory of Human Anatomy and Embryology, UA RUCA, B-2020 Antwerpen, Belgium.
J Morphol. 2003 Sep;257(3):364-74. doi: 10.1002/jmor.10123.
When in unincubated chicken blastoderms the Rauber's sickle is (sub)totally mechanically removed by selective scraping, the further evolution of the blastoderm in culture is often profoundly disturbed, going from only expansion of the upper layer and preneural plate formation to the development of a slowly growing miniature embryo. Our results suggest that the developmental potencies of the embryo are related to the presence or absence of Rauber's sickle material left after its removal. This can be checked after culture by the presence or nonpresence of junctional endoblast (derived from Rauber's sickle) and the concomitant induction of blood islands in the immediate neighborhood. Our study thus indicates that without Rauber's sickle (in the cases of successful total selective removal), an avian blastoderm cannot develop normally, even in the presence of an intact caudal marginal zone. After placing a fragment of quail sickle endoblast on the anti-sickle region of unincubated chicken blastoderms from which the Rauber's sickle was (sub)totally removed, different developmental scenarios were seen, according to the degree of removal, both in the anti-sickle as in the sickle regions. 1) If Rauber's sickle activity is strongly reduced, then besides a centripetally directed miniature embryo, induced by the remnants of the autochthonous Rauber's sickle, an additional centripetally directed embryo or preneural plate (without accompanying blood islands) develops in the anti-sickle region under inductory influence of the apposed quail sickle endoblast. We make a distinction between a neural plate and a preneural plate. The latter consists of a thickening of the upper layer (with the same initial aspect as a neural plate) adjacent to endophyll or sickle endoblast in the absence of chordomesoblast and gastrulation phenomena. 2) If Rauber's sickle activity is totally absent, then the inducing power of the sickle endoblast fragment becomes maximal and, starting from the anti-sickle region, one single embryo (without blood islands) extending over the whole area centralis appears. 3) If much of the Rauber's sickle material has been left in the blastoderm, then the inducing activity of the sickle endoblast, placed on the anti-sickle region, will be totally suppressed (although the sickle endoblast remains intact) and neither a preneural plate nor a primitive streak was induced. After placing a fragment of quail sickle endoblast on the anti-sickle region of an unincubated chicken blastoderm from which the Rauber's sickle and surrounding tissues were completely excised, an embryo was always induced by the sickle endoblast in the adjacent upper layer of this anti-sickle region. In the absence of sickle endoblast, this never occurred. Thus, our experiments demonstrate that in the absence of the Rauber's sickle, a parent tissue (sickle endoblast) induces both gastrulation and neurulation phenomena, while in the full presence of Rauber's sickle these functions are totally suppressed. Moreover, Rauber's sickle not only organizes gastrulation and blood island formation by itself but also influences neurulation at a distance (in space and time) by part of its cell lineage (i.e., sickle endoblast). Our study suggests that the inhibitory effect of Rauber's sickle on its parent tissue (sickle endoblast) represents an early mechanism impairing polyembryony, so that only a single primary major organizer (Rauber's sickle) remains active in the young avian germinal disc.
在未孵化的鸡胚盘里,如果通过选择性刮除将劳伯氏镰刀(Rauber's sickle)(部分或全部)机械去除,胚盘在培养中的进一步发育常常会受到严重干扰,从仅上层扩展和神经前板形成,发展到缓慢生长的微型胚胎的发育。我们的结果表明,胚胎的发育潜能与去除劳伯氏镰刀后残留的劳伯氏镰刀物质的有无有关。这可以在培养后通过连接内胚层(源自劳伯氏镰刀)的有无以及紧邻区域血岛的伴随诱导来检查。因此,我们的研究表明,没有劳伯氏镰刀(在成功完全选择性去除的情况下),即使存在完整的尾缘区,禽胚盘也不能正常发育。在将鹌鹑镰刀内胚层片段置于未孵化的鸡胚盘的抗镰刀区域(劳伯氏镰刀已被部分或全部去除)后,根据去除程度,在抗镰刀区域和镰刀区域都出现了不同的发育情况。1)如果劳伯氏镰刀的活性大幅降低,那么除了由自身劳伯氏镰刀残余诱导产生的向心方向的微型胚胎外,在相邻鹌鹑镰刀内胚层的诱导影响下,抗镰刀区域还会发育出另一个向心方向的胚胎或神经前板(无伴随血岛)。我们区分神经板和神经前板。后者由上层增厚形成(初始外观与神经板相同),在没有脊中胚层和原肠胚形成现象的情况下,与内叶或镰刀内胚层相邻。2)如果劳伯氏镰刀的活性完全缺失,那么镰刀内胚层片段的诱导能力就会达到最大,从抗镰刀区域开始,会出现一个单一的胚胎(无血岛),扩展到整个中央区域。3)如果胚盘里残留了大量劳伯氏镰刀物质,那么置于抗镰刀区域的镰刀内胚层的诱导活性将被完全抑制(尽管镰刀内胚层保持完整),既不会诱导出神经前板,也不会诱导出原条。在将鹌鹑镰刀内胚层片段置于未孵化的鸡胚盘的抗镰刀区域(劳伯氏镰刀和周围组织已被完全切除)后,该抗镰刀区域相邻的上层中总会由镰刀内胚层诱导出一个胚胎。在没有镰刀内胚层的情况下,这种情况从未发生。因此,我们的实验表明,在没有劳伯氏镰刀的情况下,一种母体组织(镰刀内胚层)会诱导原肠胚形成和神经胚形成现象,而在劳伯氏镰刀完全存在的情况下,这些功能会被完全抑制。此外,劳伯氏镰刀不仅自身组织原肠胚形成和血岛形成,还通过其部分细胞谱系(即镰刀内胚层)在空间和时间上远距离影响神经胚形成。我们的研究表明,劳伯氏镰刀对其母体组织(镰刀内胚层)的抑制作用代表了一种早期机制,可损害多胚形成,从而使年轻禽胚盘中只有一个主要的初级组织者(劳伯氏镰刀)保持活跃。