Neurite penetration into collagen gels requires Ca2+-dependent metalloproteinase activity.
Neurite penetration into collagen gels requires Ca2+-dependent metalloproteinase activity.
复制标题
神经突渗透到胶原蛋白凝胶中需要 Ca2 依赖性金属蛋白酶活性。
DOI：
10.1159/000111884
复制
发表时间：
1989
影响因子：
2.9
通讯作者：
Williams,AG
中科院分区：
文献类型：
--
作者：
Pittman,RN;Williams,AG
培养的新生大鼠交感神经细胞释放一种钙离子依赖的金属蛋白酶，能够降解天然和变性的I型胶原。生长中轴突的生长锥体释放出很大一部分金属蛋白酶活性，提示该酶可能参与了轴突在发育过程中穿透富含胶原的细胞外基质的过程。在目前的研究中，我们研究了轴突穿透三维胶原凝胶需要钙依赖的金属蛋白酶的可能性。将交感神经元接种于胶原凝胶上，用胶原酶抑制剂HSCH_2CH[CH_2CH(CH_3)_2]-CO-Phe-Ala-NH2(HS-LFA)抑制神经元金属蛋白酶活性。HS-LFA在非常相似的浓度范围内(EC_(50)=150-200 nM)以剂量依赖的方式抑制钙依赖的金属蛋白酶活性、突起对胶原凝胶的穿透和~3H-胶原胶的释放。突起对胶原凝胶的穿透率与金属蛋白酶活性呈高度正相关(r=0.99)，与~3H-胶原凝胶释放的~3H呈高度正相关(r=0.99)。HS-LFA虽然抑制了胶原凝胶内神经突起的生长，但对胶原膜上的神经突起生长没有影响。因此，神经元钙依赖的金属蛋白酶似乎是轴突穿透和生长三维胶原基质所必需的，而不是沿着二维胶原基质生长所必需的。
Cultured sympathetic neurons from neonatal rats release a Ca2+-dependent metalloproteinase capable of degrading native and denatured type I collagen. A large fraction of the metalloproteinase activity is released from growth cones of growing neurites, which suggests that this proteinase may be involved in neurite penetration into collagen-rich extracellular matrices during development. In the present study, we investigated the possibility that neurite penetration into 3-dimensional collagen gels requires the Ca2+-dependent metalloproteinase. Sympathetic neurons were plated on top of collagen gels and the neuronal metalloproteinase activity was inhibited with the collagenase inhibitor HSCH2CH[CH2CH(CH3)2]-CO-Phe-Ala-NH2(HS-LFA). HS-LFA inhibited the Ca2+-dependent metalloproteinase activity, penetration of neurites into collagen gels and release of3H from3H-collagen gels in a dose-dependent manner over very similar concentration ranges (EC50= 150–200 nM). Highly significant correlations existed between neurite penetration into collagen gels and metalloproteinase activity (r = 0.99), and metalloproteinase activity and release of3H from3H-collagen gels (r = 0.99). Although HS-LFA inhibited the growth of neurites within collagen gels, it had no effect on neurite outgrowth on collagen films. Thus, the neuronal Ca2+-dependent metalloproteinase appears to be required for neurite penetration into and growth within 3-dimensional collagen matrices, but not for growth along a 2-dimensional collagen substrate.