Rubin B P, Dusart I, Schwab M E
Brain Research Institute, University of Zürich, Switzerland.
J Neurocytol. 1994 Apr;23(4):209-17. doi: 10.1007/BF01275525.
In previous studies two neurite growth inhibiting protein fractions of 35 and 250 kDa were identified in myelin preparations of the rat CNS. These activities were not found in the myelin of PNS. A monoclonal antibody (mAb IN-1) was raised against the 250 kDa protein fraction and selected for its ability to neutralize the inhibitory effect of CNS myelin and of both isolated protein fractions. IN-1 has been shown both in vitro and in vivo to neutralize the inhibitory effect of differentiated oligodendrocytes and CNS white matter. In the present study, the antigens of IN-1 were localized by immunohistochemistry on cryostat sections of the adult rat nervous system. The staining pattern of IN-1 was compared to that of mAbs specific for proteins found in CNS and PNS myelin. These proteins include myelin basic protein, myelin oligodendrocyte glycoprotein, and myelin associated glycoprotein. IN-1 stained white matter and myelinated fibre tracts in the CNS on sections of fresh frozen tissue fixed with 95% ethanol: 5% acetic acid (Clark's solution). Sciatic nerve myelin and spinal roots remained unstained. The staining pattern of IN-1 corresponded most closely to that of a mAb against myelin oligodendrocyte glycoprotein, a protein which occurs exclusively in CNS myelin and on differentiated oligodendrocytes.
在先前的研究中,在大鼠中枢神经系统的髓磷脂制剂中鉴定出了两种分子量分别为35 kDa和250 kDa的神经突生长抑制蛋白组分。在周围神经系统的髓磷脂中未发现这些活性。制备了一种针对250 kDa蛋白组分的单克隆抗体(mAb IN-1),并因其能够中和中枢神经系统髓磷脂以及两种分离的蛋白组分的抑制作用而被选用。IN-1已在体外和体内被证明能够中和分化的少突胶质细胞和中枢神经系统白质的抑制作用。在本研究中,通过免疫组织化学方法在成年大鼠神经系统的低温切片上定位了IN-1的抗原。将IN-1的染色模式与针对中枢神经系统和周围神经系统髓磷脂中发现的蛋白质的单克隆抗体的染色模式进行了比较。这些蛋白质包括髓磷脂碱性蛋白、髓磷脂少突胶质细胞糖蛋白和髓磷脂相关糖蛋白。在经95%乙醇:5%乙酸(克拉克溶液)固定的新鲜冷冻组织切片上,IN-1对中枢神经系统中的白质和有髓纤维束进行了染色。坐骨神经髓磷脂和脊神经根未染色。IN-1的染色模式与针对髓磷脂少突胶质细胞糖蛋白的单克隆抗体的染色模式最为相似,髓磷脂少突胶质细胞糖蛋白是一种仅存在于中枢神经系统髓磷脂和分化的少突胶质细胞上的蛋白质。