Brochu G, Maler L, Hawkes R
Department of Biochemistry, Faculty of Medicine, Laval University, Quebec, Canada.
J Comp Neurol. 1990 Jan 22;291(4):538-52. doi: 10.1002/cne.902910405.
Monoclonal antibody mab-zebrin II was generated against a crude homogenate of cerebellum and electrosensory lateral line lobe from the weakly electric fish Apteronotus leptorhynchus. On Western blots of fish cerebellar proteins, mab-zebrin II recognizes a single polypeptide antigen of apparent molecular weight 36 kD. Immunocytochemistry of apteronotid brains reveals that zebrin II immunoreactivity is confined exclusively to Purkinje cells in the corpus cerebelli, lateral valvula cerebelli, and the eminentia granularis anterior. Other Purkinje cells, in the medial valvula cerebelli and eminentia granularis posterior, are not zebrin II immunoreactive. Immunoreactive Purkinje cells are stained completely, including dendrites, axons, and somata. The antigen seems to be absent only from the nucleus. A similar distribution is seen in catfish, goldfish, and a mormyrid fish. Zebrin II immunoreactivity is also found in the rat cerebellum. Western blotting of rat cerebellar proteins reveals a single immunoreactive polypeptide, with apparent molecular weight 36 kD, as in the fish. Also as in the fish, staining in the adult rat cerebellum is confined to a subset of Purkinje cells. Peroxidase reaction product is deposited throughout the immunoreactive Purkinje cells with the exception of the nucleus. No other cells in the cerebellum express zebrin II. At higher antibody concentrations, a weak glial cross reactivity is seen in most other brain regions: we believe that this is probably nonspecific. Zebrin II+ Purkinje cells are clustered together to form roughly parasagittal bands interposed by similar nonimmunoreactive clusters. In all there are 7 zebrin II+ and 7 zebrin II- compartments in each hemicerebellum. One immunoreactive band is adjacent to the midline; two others are disposed laterally to each side in the vermis; there is a paravermal band; and finally three more bands are identified in each hemisphere. Both in number and position, these compartments correspond precisely to the bands revealed by using another antibody, mabQ113 (anti-zebrin I). In both fish and rat the compartmentation revealed by zebrin II immunocytochemistry is related to the organization of cerebellar afferent and efferent projections and may provide clues as to the fundamental architecture of the vertebrate cerebellum.
单克隆抗体mab - zebrin II是针对弱电鱼线翎电鳗的小脑和电感应侧线叶的粗匀浆产生的。在鱼小脑蛋白的蛋白质印迹法中，mab - zebrin II识别出一种表观分子量为36 kD的单一多肽抗原。线翎电鳗脑的免疫细胞化学显示，zebrin II免疫反应性仅局限于小脑本体、小脑外侧瓣和前颗粒隆起中的浦肯野细胞。小脑内侧瓣和后颗粒隆起中的其他浦肯野细胞没有zebrin II免疫反应性。免疫反应性浦肯野细胞被完全染色，包括树突、轴突和胞体。抗原似乎仅在细胞核中不存在。在鲶鱼、金鱼和一种长颌鱼中也观察到类似的分布。在大鼠小脑中也发现了zebrin II免疫反应性。大鼠小脑蛋白的蛋白质印迹法显示，与鱼一样，有一条表观分子量为36 kD的单一免疫反应性多肽。同样与鱼一样，成年大鼠小脑中的染色局限于一部分浦肯野细胞。除细胞核外，过氧化物酶反应产物沉积在整个免疫反应性浦肯野细胞中。小脑中没有其他细胞表达zebrin II。在较高抗体浓度下，在大多数其他脑区可见微弱的胶质细胞交叉反应：我们认为这可能是非特异性的。zebrin II阳性浦肯野细胞聚集在一起形成大致的矢状旁带，中间夹杂着类似的无免疫反应性簇。每个半脑中共有7个zebrin II阳性和7个zebrin II阴性区室。一条免疫反应带与中线相邻；另外两条位于蚓部两侧；有一条旁蚓带；最后在每个半球中又识别出三条带。在数量和位置上，这些区室与使用另一种抗体mabQ113（抗zebrin I）显示的带精确对应。在鱼和大鼠中，zebrin II免疫细胞化学显示的区室化与小脑传入和传出投射的组织有关，可能为脊椎动物小脑的基本结构提供线索。
