Motoike T, Sano K, Tsuneishi S, Nakamura H, Kushima Y, Hatanaka H, Takai Y
Department of Pediatrics, Kobe University School of Medicine, Japan.
Neurosci Lett. 1991 Dec 16;134(1):109-12. doi: 10.1016/0304-3940(91)90519-y.
We have studied the expression of smg p25A and synaptophysin in cultured hippocampal neurons isolated from 5-day-old rat brain by an immunocytochemical technique. In a dispersed cell culture seeded on astrocyte monolayers, well-branching neurite proliferation was observed along with age in culture. The synaptophysin immunoreactivity was present in the neuronal cell bodies and neurites at 1 and 5 days in vitro (DIV) and was eventually localized to discrete areas along neurites at 15 DIV while the immunoreactivity in cell bodies became less prominent. On the other hand, the smg p25A immunoreactivity was observed in the neuronal cell bodies and neurites at 1 through 15 DIV. The immunoreactivity for smg p25A or synaptophysin was not observed in astrocytes and this finding was confirmed by an immunoblot analysis. These results indicate that smg p25A as well as synaptophysin is present exclusively in neurons and suggest that these two synapse-associated proteins have different sites of function and different kinetics of synthesis, transport, and/or turnover in cultured hippocampal neurons.
我们采用免疫细胞化学技术，研究了从5日龄大鼠脑部分离的培养海马神经元中smg p25A和突触素的表达。在接种于星形胶质细胞单层上的分散细胞培养物中，随着培养时间的延长，观察到神经突呈良好分支并增殖。体外培养1天和5天时，神经元胞体和神经突中存在突触素免疫反应性，而在体外培养15天时，其最终定位于神经突上的离散区域，同时胞体中的免疫反应性变得不那么明显。另一方面，在体外培养1至15天期间，在神经元胞体和神经突中均观察到smg p25A免疫反应性。在星形胶质细胞中未观察到smg p25A或突触素的免疫反应性，免疫印迹分析证实了这一发现。这些结果表明，smg p25A以及突触素仅存在于神经元中，并表明这两种与突触相关的蛋白质在培养的海马神经元中具有不同的功能位点以及不同的合成、运输和/或周转动力学。
