Manthorpe M, Engvall E, Ruoslahti E, Longo F M, Davis G E, Varon S
J Cell Biol. 1983 Dec;97(6):1882-90. doi: 10.1083/jcb.97.6.1882.
The ability of axons to grow through tissue in vivo during development or regeneration may be regulated by the availability of specific neurite-promoting macromolecules located within the extracellular matrix. We have used tissue culture methods to examine the relative ability of various extracellular matrix components to elicit neurite outgrowth from dissociated chick embryo parasympathetic (ciliary ganglion) neurons in serum-free monolayer culture. Purified laminin from both mouse and rat sources, as well as a partially purified polyornithine-binding neurite promoting factor (PNPF-1) from rat Schwannoma cells all stimulate neurite production from these neurons. Laminin and PNPF-1 are also potent stimulators of neurite growth from cultured neurons obtained from other peripheral as well as central neural tissues, specifically avian sympathetic and sensory ganglia and spinal cord, optic tectum, neural retina, and telencephalon, as well as from sensory ganglia of the neonatal mouse and hippocampal, septal, and striatal tissues of the fetal rat. A quantitative in vitro bioassay method using ciliary neurons was used to (a) measure and compare the specific neurite-promoting activities of these agents, (b) confirm that during the purification of laminin, the neurite-promoting activity co-purifies with the laminin protein, and (c) compare the influences of antilaminin antibodies on the neurite-promoting activity of laminin and PNPF-1. We conclude that laminin and PNPF-1 are distinct macromolecules capable of expressing their neurite-promoting activities even when presented in nanogram amounts. This neurite-promoting bioassay currently represents the most sensitive test for the biological activity of laminin.
在发育或再生过程中，轴突在体内穿过组织生长的能力可能受细胞外基质中特定神经突促进大分子可用性的调节。我们利用组织培养方法，在无血清单层培养中，检测各种细胞外基质成分从解离的鸡胚副交感神经（睫状神经节）神经元引发神经突生长的相对能力。从小鼠和大鼠来源纯化的层粘连蛋白，以及从大鼠雪旺细胞瘤细胞中部分纯化的聚鸟氨酸结合神经突促进因子（PNPF-1），均能刺激这些神经元产生神经突。层粘连蛋白和PNPF-1也是从其他外周以及中枢神经组织获得的培养神经元神经突生长的有效刺激物，特别是禽类交感神经节和感觉神经节、脊髓、视顶盖、神经视网膜和端脑，以及新生小鼠的感觉神经节和胎鼠的海马、隔区和纹状体组织。使用睫状神经元的定量体外生物测定方法用于：（a）测量和比较这些因子的特定神经突促进活性；（b）确认在层粘连蛋白纯化过程中，神经突促进活性与层粘连蛋白蛋白共纯化；（c）比较抗层粘连蛋白抗体对层粘连蛋白和PNPF-1神经突促进活性的影响。我们得出结论，层粘连蛋白和PNPF-1是不同的大分子，即使以纳克量呈现也能表达其神经突促进活性。这种神经突促进生物测定目前是层粘连蛋白生物活性最敏感的测试。
