Schinstine M, Cornbrooks C J
Department of Anatomy and Neurobiology, University of Vermont College of Medicine, Burlington 05405.
Brain Res. 1988 Sep 1;471(1):23-37. doi: 10.1016/0165-3806(88)90150-2.
In the present study, we have examined the growth characteristics of CNS neurons on type I collagen, detergent-treated collagen (dColl), Schwann cell-derived basal lamina (SC-BL), and purified laminin substrata. Neurons from the cerebral cortex, septal basal forebrain, and lumbosacral spinal cord were obtained from embryonic age (E) 15 and E18 rats and grown in vitro as explants on the test substrata. Neurons from either embryonic age displayed radial neurite outgrowth on collagen and dColl substrata. However, pretreatment of collagen with detergents slightly diminished its ability to support neurite outgrowth, as evidence by the 20-40% decrease in the rate of neurite growth on dColl versus the rate calculated for neurons on collagen. In contrast to the similar growth characteristics of E15 and E18 neurons on collagen and dColl, the pattern of neurite outgrowth for CNS neurons on SC-BL and laminin substrata was age dependent. Most E15 neurons grown on SC-BL extended neurites that grew identically to those observed on dColl; these 'non-orienting' neurites maintained a radial orientation to their outgrowth despite encountering interposing channels of SC-BL and grew at rates equal to that calculated for neurons on dColl. E15 neurons placed on laminin substrata showed similar growth patterns and rates equal to that calculated for neurons on dColl. E15 neurons placed on laminin substrata showed similar growth patterns and rates to neurons on collagen. In contrast, neurons from E18 rats exhibited neurites that preferentially grew in intimate association with SC-BL channels once contact with the channels was established. These 'orienting' neurites faithfully elongated within the SC-BL and demonstrated a 1.4- to 2.0-fold increase in growth rate compared with the sister cultures of neurons grown on dColl. Furthermore, E18 neurons exhibited a 1.4-fold increase in growth on laminin compared with E18 neurons grown on collagen. A minor population of neurites exhibiting similar characteristics to orienting neurites was also observed in E15 cultures. It is hypothesized that orienting and non-orienting neurites reflect the outgrowth of 'regenerating' and 'developing' neurons, respectively, and may indicate an inherent difference in the ability of regenerating and developing neurons to recognize and respond to the same guidance signals.
在本研究中,我们检测了中枢神经系统(CNS)神经元在I型胶原蛋白、经去污剂处理的胶原蛋白(dColl)、雪旺细胞衍生的基底膜(SC - BL)和纯化层粘连蛋白基质上的生长特性。来自胚胎期(E)15和E18大鼠的大脑皮质、隔区基底前脑和腰骶脊髓的神经元,作为外植体在测试基质上进行体外培养。来自任一胚胎期的神经元在胶原蛋白和dColl基质上均表现出放射状神经突生长。然而,用去污剂预处理胶原蛋白会略微降低其支持神经突生长的能力,dColl上神经突生长速率比胶原蛋白上神经元的计算速率降低20 - 40%,这证明了这一点。与E15和E18神经元在胶原蛋白和dColl上相似的生长特性相反,CNS神经元在SC - BL和层粘连蛋白基质上的神经突生长模式具有年龄依赖性。大多数在SC - BL上生长的E15神经元伸出的神经突生长方式与在dColl上观察到的相同;这些“非定向”神经突在生长过程中尽管遇到SC - BL的间隔通道,仍保持放射状生长方向,且生长速率与dColl上神经元的计算速率相同。置于层粘连蛋白基质上的E15神经元表现出相似的生长模式,生长速率与胶原蛋白上的神经元相同。相比之下,一旦与通道建立接触,来自E18大鼠的神经元的神经突会优先与SC - BL通道紧密结合生长。这些“定向”神经突在SC - BL内忠实地伸长,与在dColl上生长的神经元姐妹培养物相比,生长速率增加了1.4至2.0倍。此外,与在胶原蛋白上生长的E18神经元相比,E18神经元在层粘连蛋白上的生长增加了1.4倍。在E15培养物中也观察到一小部分神经突表现出与定向神经突相似的特征。据推测,定向和非定向神经突分别反映了“再生”和“发育”神经元的生长,可能表明再生和发育神经元识别和响应相同引导信号的能力存在内在差异。