Hatten M E, Liem R K, Mason C A
J Cell Biol. 1984 Jan;98(1):193-204. doi: 10.1083/jcb.98.1.193.
Specific interactions between neurons and glia dissociated from early postnatal mouse cerebellar tissue were studied in vitro by indirect immunocytochemical staining with antisera raised against purified glial filament protein, galactocerebroside, and the NILE glycoprotein. Two forms of cells were stained with antisera raised against purified glial filament protein. The first, characterized by a cell body 9 microns diam and processes 130-150 microns long, usually had two to three neurons associated with them and resembled Bergmann glia. The second had a slightly larger cell body with markedly shorter arms among which were nestled several dozen neuronal cells, and resembled astrocytes of the granular layer. Staining with monoclonal antisera raised against purified galactocerebroside revealed the presence of immature oligodendroglia in the cultures. These glial cells constituted approximately 2% of the total cell population in the cultures and, in contrast to astroglia, did not form specific contacts with neurons. Staining with two neuronal markers, antisera raised against purified NILE glycoprotein and tetanus toxin, revealed that most cells associated with presumed astroglia were small neurons (5-8 microns). After 1-2 d in culture, some stained neurons had very fine, short processes. Nearly all of the processes greater than 10-20 micron long were glial in origin. Electron microscopy also demonstrated the presence of two forms of astroglia in the cultures, each with a different organizing influence on cerebellar neurons. Most neurons associated with astroglia were granule neurons, although a few larger neurons sometimes associated with them. Time-lapse video microscopy revealed extensive cell migration (approximately 10 microns/h) along the arms of Bergmann-like astroglia. In contrast, cells did not migrate along the arms of astrocyte-like astroglia, but remained stationary at or near branch points. Growth cone activity, pulsating movements of cell perikarya, and ruffling of the membranes of glial and neuronal processes were also seen.
利用针对纯化的神经胶质纤维酸性蛋白、半乳糖脑苷脂和尼罗糖蛋白制备的抗血清，通过间接免疫细胞化学染色，对出生后早期小鼠小脑组织解离出的神经元和神经胶质细胞之间的特异性相互作用进行了体外研究。用针对纯化神经胶质纤维酸性蛋白制备的抗血清对两种细胞进行了染色。第一种细胞的特征是细胞体直径为9微米，突起长130 - 150微米，通常有两到三个神经元与之相关联，类似于伯格曼神经胶质细胞。第二种细胞的细胞体稍大，臂明显较短，其中簇拥着几十个体神经元细胞，类似于颗粒层的星形胶质细胞。用针对纯化半乳糖脑苷脂制备的单克隆抗血清染色显示培养物中存在未成熟的少突胶质细胞。这些神经胶质细胞约占培养物中细胞总数的2%，与星形胶质细胞不同，它们不与神经元形成特异性接触。用两种神经元标记物，即针对纯化尼罗糖蛋白和破伤风毒素制备的抗血清染色显示，与假定的星形胶质细胞相关联的大多数细胞是小神经元（5 - 8微米）。培养1 - 2天后，一些染色的神经元有非常细短的突起。几乎所有长度大于10 - 20微米的突起都起源于神经胶质细胞。电子显微镜也显示培养物中存在两种形式的星形胶质细胞，每种对小脑神经元都有不同的组织影响。与星形胶质细胞相关联的大多数神经元是颗粒神经元，尽管有时也有一些较大的神经元与之相关联。延时视频显微镜显示细胞沿着类伯格曼星形胶质细胞的臂进行广泛迁移（约10微米/小时）。相比之下，细胞不沿着类星形胶质细胞的臂迁移，而是在分支点处或附近保持静止。还观察到生长锥活动、细胞周缘的脉动运动以及神经胶质细胞和神经元突起膜的皱褶。
