Liu Haiping, Ng Cherry E L, Tang Bor Luen
NCA laboratory, Institute of Molecular and Cell Biology, 30 Medical Drive, Singapore, Singapore.
Neurosci Lett. 2002 Aug 16;328(3):257-60. doi: 10.1016/s0304-3940(02)00528-1.
The longest central nervous system (CNS) specific isoform of the major myelin-associated inhibitor of neurite growth, Nogo-A, has previously been shown to be expressed largely in oligodendrocytes. Using an antibody raised against a recombinant fusion protein comprising amino acids 223-399 of Nogo-A, we show in this report that Nogo-A is also expressed in the cell body of a distinct set of CNS neurons. The antibody detects a single protein band of 220 kDa in rat brain lysate and neuroblastoma cell lysates. Immunofluorescent analyses reveal that Nogo-A is found largely in the endoplasmic reticulum of neuroblastoma cell lines SH-SY5Y and NIE-115. In the mouse CNS, Nogo-A can be found in specific subsets of neuronal cell bodies in addition to oligodendrocytes, but not glial fibrillary associated protein positive astrocytes or MAC-1 positive microglia. Our results provide a conclusive demonstration of the expression of Nogo-A in CNS neurons, which suggests that Nogo-A may have distinct endogenous roles in neurons other than its known ability to inhibit neurite growth.
神经突生长的主要髓鞘相关抑制剂Nogo-A最长的中枢神经系统(CNS)特异性同工型,此前已被证明主要在少突胶质细胞中表达。在本报告中,我们使用针对包含Nogo-A第223 - 399位氨基酸的重组融合蛋白产生的抗体,证明Nogo-A也在一组独特的CNS神经元的细胞体中表达。该抗体在大鼠脑裂解物和神经母细胞瘤细胞裂解物中检测到一条220 kDa的单一蛋白条带。免疫荧光分析显示,Nogo-A主要存在于神经母细胞瘤细胞系SH-SY5Y和NIE-115的内质网中。在小鼠中枢神经系统中,除少突胶质细胞外,在神经元细胞体的特定亚群中也可发现Nogo-A,但在胶质纤维酸性蛋白阳性星形胶质细胞或MAC-1阳性小胶质细胞中未发现。我们的结果确凿地证明了Nogo-A在中枢神经系统神经元中的表达,这表明Nogo-A除了其已知的抑制神经突生长的能力外,可能在神经元中具有独特的内源性作用。