Pesacreta T C, Byers T J, Dubreuil R, Kiehart D P, Branton D
Department of Cellular and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.
J Cell Biol. 1989 May;108(5):1697-709. doi: 10.1083/jcb.108.5.1697.
The distribution of alpha-spectrin in Drosophila embryos was determined by immunofluorescence using affinity-purified polyclonal or monoclonal antibodies. During early development, spectrin is concentrated near the inner surface of the plasma membrane, in cytoplasmic islands around the syncytial nuclei, and, at lower concentrations, throughout the remainder of the cytoplasm of preblastoderm embryos. As embryogenesis proceeds, the distribution of spectrin shifts with the migrating nuclei toward the embryo surface so that, by nuclear cycle 9, a larger proportion of the spectrin is concentrated near the plasma membrane. During nuclear cycles 9 and 10, as the nuclei reach the cell surface, the plasma membrane-associated spectrin becomes concentrated into caps above the somatic nuclei. Concurrent with the mitotic events of the syncytial blastoderm period, the spectrin caps elongate at interphase and prophase, and divide as metaphase and anaphase progress. During cellularization, the regions of spectrin concentration appear to shift: spectrin increases near the growing furrow canal and concomitantly increases at the embryo surface. In the final phase of furrow growth, the shift in spectrin concentration is reversed: spectrin decreases near the furrow canal and concomitantly increases at the embryo surface. In gastrulae, spectrin accumulates near the embryo surface, especially at the forming amnioproctodeal invagination and cephalic furrow. During the germband elongation stage, the total amount of spectrin in the embryo increases significantly and becomes uniformly distributed at the plasma membrane of almost all cell types. The highest levels of spectrin are in the respiratory tract cells; the lowest levels are in parts of the forming gut. The spatial and temporal changes in spectrin localization suggest that this protein plays a role in stabilizing rather than initiating changes in structural organization in the embryo.
使用亲和纯化的多克隆或单克隆抗体,通过免疫荧光法确定了果蝇胚胎中α-血影蛋白的分布。在早期发育过程中,血影蛋白集中在质膜内表面附近、合胞体细胞核周围的细胞质岛中,并且在胚盘形成前胚胎的其余细胞质中浓度较低。随着胚胎发生的进行,血影蛋白的分布随着迁移的细胞核向胚胎表面移动,因此到核周期9时,更大比例的血影蛋白集中在质膜附近。在核周期9和10期间,当细胞核到达细胞表面时,与质膜相关的血影蛋白会集中到体细胞核上方的帽状结构中。在合胞体胚盘期的有丝分裂事件同时发生时,血影蛋白帽在间期和前期伸长,并在中期和后期进展时分裂。在细胞化过程中,血影蛋白浓度区域似乎发生了变化:血影蛋白在生长的沟道附近增加,并在胚胎表面同时增加。在沟道生长的最后阶段,血影蛋白浓度的变化方向相反:血影蛋白在沟道附近减少,并在胚胎表面同时增加。在原肠胚中,血影蛋白在胚胎表面附近积累,特别是在形成羊膜肛凹和头沟处。在胚带伸长阶段,胚胎中血影蛋白的总量显著增加,并在几乎所有细胞类型的质膜上均匀分布。血影蛋白水平最高的是呼吸道细胞;最低的是正在形成的肠道部分。血影蛋白定位的时空变化表明,这种蛋白质在稳定胚胎结构组织变化而非启动变化方面发挥作用。