Beer Justinus, Technau Gerhard M, Campos-Ortega Jose A
Institut for Entwicklungsphysiologie, Universität zu Köln, Gyrhofstraße 17, D-5000, Köln 41, Germany.
Rouxs Arch Dev Biol. 1987 Apr;196(4):222-230. doi: 10.1007/BF00376346.
We describe the results of cell transplantation experiments performed to investigate mesodermal lineages in Drosophila melanogaster, particularly the lineages of the somatic muscles, the visceral muscles and the fat body. Cells to be transplanted were labelled by injecting a mixture of horseradish peroxidase (HRP) and fluorescein-dextran (FITC) in wild-type embryos at the syncytial blastoderm stage. For transplantation cells were removed from the ventral furrow, 8-12 min after the start of gastrulation, and individually transplanted into homotopic or heterotopic locations of unlabelled wild-type hosts of the same age. HRP labelling in the resulting cell clones was demonstrated histochemically in the fully developed embryo; histotypes could be distinguished without ambiguity. Mesodermal cells were already found to be committed to mesodermal fates at the time of transplantation. They developed only into mesodermal derivatives and did not integrate in non-mesodermal organs upon heterotopical transplantation. No evidence was found for commitment to any particular mesodermal organ at the time of transplantation. The majority of somatic muscle clones contributed cells to only one segment. However, clones were not infrequently distributed through two or even three segments. Clones of fat body cells were generally restricted to a small region. However, cells of clones of visceral musculature were widely distributed. With respect to the proliferative abilities of transplanted cells the clones were difficult to interpret due to the syncytial character of the somatic musculature and the fact that the organization of the other organs is poorly understood. Evidence from histological observations of developing normal embryos indicates only three mitoses for mesodermal cells. Clones larger than seven cells were not found when embryos were fixed previous to germ-band shortening; larger clones were found in the fat body and visceral musculature after fixing the embryos at the end of organogenesis. Quantitative considerations suggest that a few mesodermal cells might perform more than three mitoses.
我们描述了为研究黑腹果蝇中胚层谱系,特别是体壁肌肉、内脏肌肉和脂肪体的谱系而进行的细胞移植实验结果。在合胞体胚盘阶段,通过向野生型胚胎中注射辣根过氧化物酶(HRP)和荧光素葡聚糖(FITC)的混合物来标记待移植的细胞。对于移植,在原肠胚形成开始后8 - 12分钟从腹沟中取出细胞,并将其单独移植到相同年龄的未标记野生型宿主的同位或异位位置。在完全发育的胚胎中通过组织化学方法显示所得细胞克隆中的HRP标记;组织类型可以明确区分。在移植时已发现中胚层细胞已确定为中胚层命运。它们仅发育为中胚层衍生物,在异位移植时不整合到非中胚层器官中。在移植时未发现有确定为任何特定中胚层器官的证据。大多数体壁肌肉克隆仅向一个节段贡献细胞。然而,克隆也经常分布在两个甚至三个节段中。脂肪体细胞克隆通常局限于一个小区域。然而,内脏肌肉组织克隆的细胞分布广泛。关于移植细胞的增殖能力,由于体壁肌肉组织的合胞体特性以及其他器官的组织结构了解不足,这些克隆难以解释。对正常发育胚胎的组织学观察证据表明中胚层细胞仅进行三次有丝分裂。在胚带缩短之前固定胚胎时未发现大于七个细胞的克隆;在器官发生结束时固定胚胎后,在脂肪体和内脏肌肉组织中发现了更大的克隆。定量分析表明,一些中胚层细胞可能进行超过三次有丝分裂。