Hertzler P L, Wang S W, Clark W H
Department of Zoology, University of California, Davis 95616.
Dev Biol. 1994 Aug;164(2):333-44. doi: 10.1006/dbio.1994.1204.
The fate map of 2- and 4-cell-stage Sicyonia ingentis embryos was determined by microinjection of lysyl-rhodamine-dextran into single blastomeres. Microinjected embryos were cultured to the limb bud stage, when the body plan of the nauplius larva was evident. The animal blastomere, AB, gave rise to anterior ectoderm, while the vegetal blastomere, CD, gave rise to posterior structures, including the invagination site during gastrulation. The A blastomere gave rise to mirror-image patterns of dorsal-lateral ectoderm, while the B blastomere gave rise to anterior, ventral ectoderm. The C blastomere gave rise to posterior, dorsal-lateral ectoderm, complementary to the A pattern, as well as some naupliar mesoderm. The D blastomere gave rise to mesendoderm, naupliar mesoderm, and some posterior ectoderm. To study the specification of the early blastomeres, they were microsurgically separated and cultured in isolation. Two mesendoderm cells formed in 1/2, 1/4, 1/8, and 1/16 blastomeres in embryos dissociated at the 2-, 4-, 8-, and 16-cell stages, respectively. CD and D blastomeres could be distinguished by their larger size and gave rise to the mesendoderm cells. Archenteron formation and elongation of the embryo occurred in CD but not in AB isolates. Isolated blastomeres were recombined in various ways to determine whether their state of commitment could be altered in different cellular environments. Duplicated mesendoderm cells and archenterons formed in CD + CD recombinations, while AB + AB recombinations formed blastulae but did not produce mesendoderm cells and did not invaginate. The normal number of mesendoderm cells and a single archenteron formed in D + AB recombinations, while C + AB recombinations remained as blastulae and did not form mesendoderm cells. The results suggest that the mesendoderm cells are autonomously specified, possibly by cytoplasmic localization at the vegetal pole. The mesendoderm may also function as a signaling region to organize other developmental events.
通过将赖氨酰罗丹明 - 葡聚糖显微注射到单个卵裂球中,确定了2细胞期和4细胞期的大型西鲱(Sicyonia ingentis)胚胎的命运图谱。显微注射后的胚胎被培养至肢芽期,此时无节幼虫的身体结构已清晰可见。动物极卵裂球AB产生前外胚层,而植物极卵裂球CD产生后结构,包括原肠胚形成期间的内陷部位。A卵裂球产生背外侧外胚层的镜像模式,而B卵裂球产生前腹侧外胚层。C卵裂球产生后背外侧外胚层,与A模式互补,以及一些无节幼虫中胚层。D卵裂球产生中内胚层、无节幼虫中胚层和一些后外胚层。为了研究早期卵裂球的特化,将它们显微手术分离并单独培养。在分别于2细胞期、4细胞期、8细胞期和16细胞期解离的胚胎中,1/2、1/4、1/8和1/16卵裂球中形成了两个中内胚层细胞。CD和D卵裂球可通过其较大的尺寸区分,并产生中内胚层细胞。原肠形成和胚胎伸长发生在CD分离物中,而不是AB分离物中。分离的卵裂球以各种方式重新组合,以确定它们的特化状态在不同的细胞环境中是否可以改变。在CD + CD重组中形成了重复的中内胚层细胞和原肠,而AB + AB重组形成囊胚,但不产生中内胚层细胞,也不内陷。在D + AB重组中形成了正常数量的中内胚层细胞和单个原肠,而C + AB重组则保持为囊胚,不形成中内胚层细胞。结果表明,中内胚层细胞可能通过植物极的细胞质定位自主特化。中内胚层也可能作为一个信号区域来组织其他发育事件。