DeGeorge J J, Slepecky N, Carbonetto S
Dev Biol. 1985 Oct;111(2):335-51. doi: 10.1016/0012-1606(85)90488-9.
Dorsal root ganglion neurons in culture proceed through a series of shape changes before growing nerve fibers. These shape changes involve: attachment to the substratum, extension of filopodia, and spreading of part of the cell to form broad lamellipodia. With the formation of lamellipodia, neurons adhere firmly to the substratum and retrogradely transport lectins (concanavalin A, wheat germ agglutinin) on their surfaces. In unspread neurons concanavalin A, but not wheat germ agglutinin, rapidly stimulates lamellipodium formation and neuron-substratum adhesion. Neurons treated with concanavalin A also have more, branched nerve fibers than untreated neurons, but otherwise appear similar. These effects of concanavalin A are concentration dependent, blocked by alpha-methyl-D-mannoside (100 mM), and are accompanied by receptor redistribution. Stimulation of lamellipodium extension by concanavalin A is inhibited by low temperature (4 degrees C), 2,4-dinitrophenol (0.2 mM), cytochalasin D (4 microM), or trifluoperazine (10 microM), but not by cycloheximide (360 microM) or colchicine (12.5 microM). Attachment of neurons to the culture substratum was affected little by these treatments. These results indicate differences in the neuron's metabolic requirements for simple attachment to the substratum and the early phases of nerve fiber growth. Moreover, they suggest a convenient system in which to study the cellular and biochemical events of rapid nerve fiber outgrowth in primary neuronal cultures.
培养中的背根神经节神经元在长出神经纤维之前会经历一系列形态变化。这些形态变化包括:附着于基质、丝状伪足的伸展以及细胞部分区域展开形成宽的片状伪足。随着片状伪足的形成,神经元牢固地附着于基质,并在其表面逆行运输凝集素(刀豆球蛋白A、麦胚凝集素)。在未展开的神经元中,刀豆球蛋白A而非麦胚凝集素能迅速刺激片状伪足形成和神经元 - 基质黏附。用刀豆球蛋白A处理的神经元也比未处理的神经元有更多且分支的神经纤维,但在其他方面看起来相似。刀豆球蛋白A的这些作用呈浓度依赖性,可被α - 甲基 - D - 甘露糖苷(100 mM)阻断,并伴有受体重新分布。刀豆球蛋白A对片状伪足伸展的刺激可被低温(4℃)、2,4 - 二硝基苯酚(0.2 mM)、细胞松弛素D(4 μM)或三氟拉嗪(10 μM)抑制,但不受放线菌酮(360 μM)或秋水仙碱(12.5 μM)抑制。这些处理对神经元附着于培养基质的影响很小。这些结果表明神经元在简单附着于基质和神经纤维生长早期阶段的代谢需求存在差异。此外,它们提示了一个方便的系统,可用于研究原代神经元培养中神经纤维快速长出的细胞和生化事件。