Schlaepfer W W, Lee C, Trojanowski J Q, Lee V M
J Neurochem. 1984 Sep;43(3):857-64. doi: 10.1111/j.1471-4159.1984.tb12809.x.
Alterations occurring in nerve proteins of transected nerves were studied in rat sciatic nerves using polyclonal and monoclonal antibodies to identify and monitor neurofilament (NF) epitopes among nerve proteins following their electrophoresis and transfer to nitrocellulose paper. Immunoblot methods identified NF epitopes in NF triplet proteins (Mr 200,000, 150,000, and 68,000) and in NF nontriplet proteins (all other immunobands below Mr 200,000 and above Mr 40,000). NF triplet and nontriplet proteins were Triton-insoluble in both untransected and transected nerves. Extensive loss of NF triplet and most nontriplet proteins occurred during the 24-48-h period following nerve transection and was attributed to proteolytic degradation. Loss of protease-labile NF proteins led to a markedly reduced level of NF immunoreactivity in 2-day transected nerve. NF proteins which survived the 2-day posttransectional period were considered to represent protease-stable NF fragments. These fragments persisted in transected nerve for periods of at least 35 days. Most protease-stable NF fragments which retained immunoreactivity had Mr of 57,000-65,000. Low concentrations of the same immunobands were present in untransected nerves.
利用多克隆抗体和单克隆抗体，通过电泳和转印至硝酸纤维素膜后识别和监测神经蛋白中的神经丝（NF）表位，对大鼠坐骨神经横断后神经蛋白发生的变化进行了研究。免疫印迹法在NF三联体蛋白（分子量200,000、150,000和68,000）以及NF非三联体蛋白（分子量低于200,000且高于40,000的所有其他免疫条带）中识别出NF表位。在未横断和横断的神经中，NF三联体和非三联体蛋白均不溶于 Triton。神经横断后24 - 48小时内，NF三联体和大多数非三联体蛋白大量丢失，这归因于蛋白水解降解。蛋白酶敏感的NF蛋白丢失导致横断2天的神经中NF免疫反应性水平显著降低。横断后2天仍存活的NF蛋白被认为代表蛋白酶稳定的NF片段。这些片段在横断神经中持续存在至少35天。大多数保留免疫反应性的蛋白酶稳定NF片段分子量为57,000 - 65,000。在未横断的神经中也存在低浓度的相同免疫条带。
