CHONDROITIN SULFATE PROTEOGLYCANS IN THE RAT-BRAIN - CANDIDATES FOR AXON BARRIERS OF SENSORY NEURONS AND THE POSSIBLE MODIFICATION BY LAMININ OF THEIR ACTIONS
CHONDROITIN SULFATE PROTEOGLYCANS IN THE RAT-BRAIN - CANDIDATES FOR AXON BARRIERS OF SENSORY NEURONS AND THE POSSIBLE MODIFICATION BY LAMININ OF THEIR ACTIONS
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DOI：
10.1111/j.1460-9568.1995.tb00665.x
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发表时间：
1995-04-01
影响因子：
3.4
通讯作者：
OOHIRA, A
中科院分区：
文献类型：
--
作者：
KATOHSEMBA, R;MATSUDA, M;OOHIRA, A
将从大鼠脑中纯化的硫酸软骨素蛋白聚糖（CSPGs）添加到PC12D细胞的培养基中，可以抑制PC12D细胞的增殖和神经突的生长。因此，我们研究了几种细胞外成分对CSPGs对PC12D细胞抑制作用的影响，以及它们在大鼠胚胎中的免疫细胞化学分布，以确定体外研究结果是否可以在体内复制。脑CSPGs对PC12D细胞的抑制作用需要在基质上涂上聚赖氨酸。用层粘连蛋白或纤维连接蛋白对聚赖氨酸包被的培养皿进行预处理，可促进PC12D细胞的神经突生长。层粘连蛋白和纤维连接蛋白，而不是胶原（I型和IV型）和cell - tak（细胞粘附分子），以浓度依赖的方式阻止脑CSPGs的抑制作用。层粘连蛋白（或纤维连接蛋白）使CSPG效应减少50%的剂量对神经突生长的影响为1.5（或25）μ g/ml，对增殖的影响为2.2（或28）μ g/ml。碟载CSPGs与层粘连蛋白减少50%所需的比例约为50:1 （wt/wt）。任何来源的层粘连蛋白都有相同的效果。脑CSPGs还能明显抑制背根神经节外植体和原代培养背根神经节神经元纤维的生长。在妊娠第13.5天，当DRG神经元轴突向神经管延伸时，在大鼠胚胎的边界帽和顶板中检测到神经can（大脑中的主要CSPG）样免疫反应性。层粘连蛋白（CSPGs）和腱蛋白（tenascin）的分布分别与神经蛋白（neurocan）有细微或显著的差异。这些结果表明，脑CSPGs可以对来自背根神经节的轴突生长产生障碍，CSPGs与层粘连蛋白的比例可能在调节轴突生长中起重要作用。
The addition of chondroitin sulphate proteoglycans (CSPGs), purified from the rat brain, to the culture medium of PC12D cells inhibited their proliferation and neurite outgrowth. Therefore, we investigated the effects of several extracellular components on the inhibitory actions of CSPGs on PC12D cells, as well as their immunocytochemical distribution in the rat embryo to determine whether the findings in vitro could be reproduced in vivo, Coating of the substratum with polylysine was necessary for the appearance of the inhibitory effects of brain CSPGs on PC12D cells. The additional pretreatment of polylysine-coated dishes with laminin or fibronectin promoted the outgrowth of neurites from PC12D cells. Laminin and fibronectin, but not collagen (types I and IV) and CELL-TAK (cell adhesion molecules), prevented the inhibitory effects of brain CSPGs in a concentration-dependent manner. Doses producing 50% reduction by laminin (or fibronectin) of the CSPG effects were 1.5 (or 25) mu g/ml for neurite outgrowth and 2.2 (or 28) mu g/ml for proliferation. The ratio of dish-attached CSPGs to laminin necessary for 50% reduction was about similar to 50:1 (wt/wt). Laminin from any source had the same effect. Brain CSPGs also obviously impeded the growth of fibres from dorsal root ganglion explants and primary cultured dorsal root ganglion neurons. Neurocan (a major CSPG in the brain)-like immunoreactivity was detected in the boundary caps and roof plate in the rat embryo at 13.5 days of gestation, when DRG neurons were extending their axons to the neural tube. The distributions of laminin and tenascin appeared, respectively, to be slightly and considerably different from that of neurocan, These results suggest that brain CSPGs can generate barriers to the growth of axons from the dorsal root ganglion and that the ratio of CSPGs to laminin may be important in regulating such growth.