Phosphacan and neurocan are repulsive substrata for adhesion and neurite extension of adult rat dorsal root ganglion neurons in vitro
Phosphacan and neurocan are repulsive substrata for adhesion and neurite extension of adult rat dorsal root ganglion neurons in vitro
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DOI：
10.1016/s0014-4886(03)00090-6
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发表时间：
2003-07-01
影响因子：
5.3
通讯作者：
Kawano, H
中科院分区：
文献类型：
--
作者：
Sango, K;Oohira, A;Kawano, H
磷酸蛋白聚糖（PC）和神经蛋白聚糖（NC）是神经组织中主要的硫酸软骨素蛋白聚糖（CS-PG），参与神经发育和再生过程中细胞粘附和神经突生长的调节。在本研究中，我们研究了PC和NC对体外培养的成年大鼠背根神经节（DRG）神经元的附着和突起延伸的影响。用PC和NC处理聚-（L）-赖氨酸（PL）以浓度依赖性方式（10 μ g/ml > 1 μ g/ml >> 0.1 μ g/ml）显著损害神经元附着和轴突延伸，并且它们被软骨素酶ABC（ChABC）消化部分抑制。应用于培养基（1 μ g/ml）的CS-PG也显示出对神经突延伸的抑制作用，ChABC处理没有改变。这些结果表明，PC和NC的成人DRG神经元在体外的粘附和轴突再生的排斥性基质，并建议，硫酸软骨素部分和核心蛋白是负责的CS-PG的抑制作用。我们还进行了免疫组织化学分析与PC（mAb 6134）和NC（mAb 1G 2）的核心蛋白的单克隆抗体，这表明，只有少数神经元的DRG切片中被这些抗体染色。与此相反，大多数DRG神经元在不同阶段（12小时，1天，2天，4天）的培养免疫反应mAb 6134和mAb 1G 2。将这些发现结合在一起，在培养的神经元中表达的CS-PG可能在附着、存活和轴突再生的调节中起作用。(C)2003 Elsevier Science（美国）。All rights reserved.
Phosphacan (PC) and neurocan (NC) are major chondroitin sulfate proteoglycans (CS-PGs) in nervous tissue and are involved in the modulation of cell adhesion and neurite outgrowth during neural development and regeneration. In the present study, we examined the effects of PC and NC on the attachment and neurite extension of adult rat dorsal root ganglion (DRG) neurons in vitro. Treatment with PC and NC on poly-(L)-lysine (PL) significantly impaired both neuronal attachment and neurite extension in a concentration-dependent manner (10 mug/ml > 1 mug/ml >> 0.1 mug/ml), and they were partially suppressed by chondroitinase ABC (ChABC) digestion. The CS-PGs applied to culture medium (1 mug/ml) also displayed inhibitory effects on neurite extension, which were not altered by ChABC treatment. These results show that PC and NC are repulsive substrata for adhesion and neurite regeneration of adult DRG neurons in vitro and suggest that both chondroitin sulfate moieties and core proteins are responsible for the inhibitory actions of the CS-PGs. We also conducted immunohistochemical analyses with the monoclonal antibodies to core proteins of PC (mAb 6134) and NC (mAb 1G2), which revealed that only a few neurons in the DRG section were stained with these antibodies. In contrast, most DRG neurons at different stages (12 h, 1 day, 2 days, and 4 days) in culture were immunoreactive to mAb 6134 and mAb 1G2. Taking these findings together, it is plausible that both CS-PGs expressed in the cultured neurons may play a role in the modulation of attachment, survival, and neurite regeneration. (C) 2003 Elsevier Science (USA). All rights reserved.