McKay R D, Hockfield S J
Proc Natl Acad Sci U S A. 1982 Nov;79(21):6747-51. doi: 10.1073/pnas.79.21.6747.
Eight hundred hybridoma lines were generated from mice immunized with the fixed gray matter of cat spinal cord. Of these lines, 47 were positive when screened immunohistochemically against sections of the cat spinal cord. Twenty-nine lines secreted antibodies that bound to neuronal antigens. Of these, 16 bound to axons only, 8 bound to axons and cell bodies, and 5 bound to cell bodies only. Eighteen lines secreted antibodies that bound to glial cells. Five lines that secreted antibodies that intensely stained spinal cord sections were cloned and screened against other parts of the central nervous system. Each of these five antibodies bound to specific subsets of neurons. For example, in the spinal cord, one antibody (Cat-301) recognized a surface determinant on the dendrites and cell bodies of neurons that, in morphology and location, resemble long-distance projection neurons. A second antibody (Cat-201) recognized an antigen in axons and in the cytoplasm of neuronal cell bodies that may be a subset of those recognized by Cat-301. A third antibody (Cat-101) recognized only axons. The subcellular localization of the antigen recognized by each antibody is the same in all areas of the central nervous system we have examined. The fact that each of the antibodies described here has a restricted distribution in the central nervous system shows that there is a high degree of molecular diversity among vertebrate neurons and that hybridoma technology can be used to explore this diversity. This class of reagents should be a useful addition to the many established techniques for studying the organization of the vertebrate central nervous system.
用猫脊髓固定灰质免疫小鼠，产生了800个杂交瘤细胞系。在这些细胞系中，用猫脊髓切片进行免疫组织化学筛选时，有47个呈阳性。29个细胞系分泌与神经元抗原结合的抗体。其中，16个仅与轴突结合，8个与轴突和细胞体结合，5个仅与细胞体结合。18个细胞系分泌与神经胶质细胞结合的抗体。对5个分泌能强烈染色脊髓切片抗体的细胞系进行克隆，并针对中枢神经系统的其他部分进行筛选。这5种抗体中的每一种都与特定的神经元亚群结合。例如，在脊髓中，一种抗体（Cat - 301）识别神经元树突和细胞体上的一种表面决定簇，这些神经元在形态和位置上类似于长距离投射神经元。第二种抗体（Cat - 201）识别轴突和神经元细胞体细胞质中的一种抗原，该抗原可能是Cat - 301识别的抗原的一个亚群。第三种抗体（Cat - 101）仅识别轴突。在所检测的中枢神经系统的所有区域中，每种抗体所识别抗原的亚细胞定位都是相同的。这里描述的每种抗体在中枢神经系统中都有受限的分布，这一事实表明脊椎动物神经元之间存在高度的分子多样性，并且杂交瘤技术可用于探索这种多样性。这类试剂应该是对许多已有的研究脊椎动物中枢神经系统组织技术的有益补充。
