Unsicker K, Skaper S D, Davis G E, Manthorpe M, Varon S
Brain Res. 1985 Jan;349(1-2):304-8. doi: 10.1016/0165-3806(85)90161-0.
We have investigated the effects of two neurite promoting factors (NPFs)--laminin and the semipurified polyornithine-binding neurite promoting factor (PNPF-1) from RN 22 Schwannoma cells--on neurite regeneration from dissociated newborn and adult rat dorsal root ganglion (DRG) neurons during 24 and 48 h culture periods in the absence of exogenous neuronotrophic factors. Both laminin and PNPF, when used to pretreat the polyornithine substratum, significantly enhanced neurite recruitment from surviving newborn and adult DRG neurons as compared to an untreated polyornithine substratum. However, the responses of newborn neurons at saturating concentrations of laminin and PNPF were consistently greater (46% neurite-bearing cells at 24 h, 81% at 48 h) than those of adult neurons (14 and 45%, respectively). The responsive neurons of both newborn and adult DRG displayed extensive neuritic networks at 48 h. The ED50 of laminin, or PNPF was 0.15-0.2 micrograms/ml for both newborn and adult neurons. The similarities in the responses of newborn and adult DRG neurons to NPFs validate the use of neurons from embryonic and newborn animals for the in vitro assays of NPFs that can be collected from injured and regenerating adult peripheral nervous tissues.
我们研究了两种神经突促进因子(NPFs)——层粘连蛋白和来自RN 22雪旺氏细胞瘤细胞的半纯化聚鸟氨酸结合神经突促进因子(PNPF-1)——在无外源性神经营养因子的情况下,对新生和成年大鼠背根神经节(DRG)解离神经元在24小时和48小时培养期内神经突再生的影响。与未处理的聚鸟氨酸基质相比,当层粘连蛋白和PNPF用于预处理聚鸟氨酸基质时,可显著增强存活的新生和成年DRG神经元的神经突募集。然而,在层粘连蛋白和PNPF饱和浓度下,新生神经元的反应(24小时时46%的神经突形成细胞,48小时时81%)始终大于成年神经元(分别为14%和45%)。在48小时时,新生和成年DRG的反应性神经元均显示出广泛的神经突网络。层粘连蛋白或PNPF对新生和成年神经元的半数有效剂量(ED50)均为0.15 - 0.2微克/毫升。新生和成年DRG神经元对NPFs反应的相似性证实了可使用来自胚胎和新生动物的神经元进行体外检测,以检测可从受伤和再生的成年周围神经组织中收集的NPFs。