ZEBRIN-II - A POLYPEPTIDE ANTIGEN EXPRESSED SELECTIVELY BY PURKINJE-CELLS REVEALS COMPARTMENTS IN RAT AND FISH CEREBELLUM
ZEBRIN-II - A POLYPEPTIDE ANTIGEN EXPRESSED SELECTIVELY BY PURKINJE-CELLS REVEALS COMPARTMENTS IN RAT AND FISH CEREBELLUM
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DOI：
10.1002/cne.902910405
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发表时间：
1990-01-22
影响因子：
2.5
通讯作者：
HAWKES, R
中科院分区：
文献类型：
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作者：
BROCHU, G;MALER, L;HAWKES, R
抗弱电鱼小脑和电感觉侧叶组织匀浆的单抗Mab-Zebrin II。在鱼类小脑蛋白的Western blotts上，Mab-Zebrin II识别一个表观分子量为36kD的单一多肽抗原。斑马素II免疫细胞化学研究表明，斑马素II免疫反应仅局限于小脑体、小脑外侧瓣和颗粒前隆起中的浦肯野细胞。小脑内侧瓣膜和后颗粒隆起中的其他浦肯野细胞不表达Zebrin II免疫反应。免疫反应阳性的Purikinje细胞完全染色，包括树突、轴突和胞体。这种抗原似乎只在细胞核中缺失。在鲶鱼、金鱼和三叉鱼中也发现了类似的分布。在大鼠小脑中也发现了Zebrin II免疫反应。大鼠小脑蛋白的Western blotting显示出单一的免疫活性多肽，其表观分子量为36kD，与鱼类中的一样。同样，与鱼类一样，成年大鼠小脑中的染色仅限于浦肯野细胞的一部分。过氧化物酶反应产物沉积在免疫反应阳性的浦肯野细胞中，但细胞核除外。小脑中没有其他细胞表达斑马素II。在抗体浓度较高的情况下，在大多数其他大脑区域可以看到弱的胶质交叉反应：我们认为这可能是非特异性的。Zebrin II+Purkinje细胞聚集在一起，形成大致的矢状面旁带，中间有类似的非免疫反应簇。每侧小脑共有7个Zebrin II+区和7个Zebrin II-区。一条免疫反应带与中线相邻；另外两条免疫反应带分布在蚯蚓两侧；存在一条卵旁带；最后，在每一半球还发现了三条免疫反应带。在数量和位置上，这些间隔与使用另一种抗体mabQ113(抗斑马素I)显示的条带精确对应。在鱼类和大鼠中，Zebrin II免疫细胞化学显示的区划与小脑传入和传出投射的组织有关，并可能为脊椎动物小脑的基本构筑提供线索。
Monoclonal antibody mab-zebrin II was generated against a crude homogenate of cerebellum and electrosensory lateral line lobe from the weakly electric fish Apteronotus leptorhynuchus. 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 Purikinje cells are stained completely, including dendrities, 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.