Shibayama-Imazu T, Okahashi I, Omata K, Nakajo S, Ochiai H, Nakai Y, Hama T, Nakamura Y, Nakaya K
Laboratory of Biological Chemistry, School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
Brain Res. 1993 Sep 17;622(1-2):17-25. doi: 10.1016/0006-8993(93)90796-p.
In the present paper, the distribution of a neuron-specific phosphoneuroprotein 14 (PNP 14) in cell and tissue was investigated in detail by the immunoblot method using affinity-purified antibody against this protein. The immunoblot of the supernatant fractions of various tissue homogenates of rat clearly demonstrated that PNP 14 was enormously rich in the brain. The content in rat brain was as much as 0.1% of the homogenate. The immunocytochemical study showed that the protein was localized at nerve endings in the cerebellum. Existence of the protein was also confirmed in cultured neuronal cells from postnatal rat midbrain, but not in glial cells. Examination of subcellular localization of PNP 14 indicates that the protein was present in synaptic plasma membranes and synaptic supernatant fractions, but not in synaptic vesicles. During the development of rat brain, PNP 14 came into existence after birth and it's amount linearly increased to a maximum at 21-28 days after birth. The content of the protein then remained at the same level for more than 10 months. We concluded that this protein is neuron specific and supposed that it may be involved in neuronal formation and function.
在本论文中,使用针对神经元特异性磷酸神经蛋白14(PNP 14)的亲和纯化抗体,通过免疫印迹法详细研究了该蛋白在细胞和组织中的分布。大鼠各种组织匀浆上清液部分的免疫印迹清楚地表明,PNP 14在脑中含量极为丰富。大鼠脑中该蛋白的含量高达匀浆的0.1%。免疫细胞化学研究表明,该蛋白定位于小脑的神经末梢。在出生后大鼠中脑的培养神经元细胞中也证实了该蛋白的存在,但在胶质细胞中未发现。对PNP 14亚细胞定位的检查表明,该蛋白存在于突触质膜和突触上清液部分,但不存在于突触小泡中。在大鼠脑发育过程中,PNP 14在出生后出现,其含量在出生后21 - 28天线性增加至最大值。然后该蛋白的含量在10多个月内保持在同一水平。我们得出结论,这种蛋白是神经元特异性的,并推测它可能参与神经元的形成和功能。