Levi G, Aloisi F, Ciotti M T, Gallo V
Brain Res. 1984 Jan 2;290(1):77-86. doi: 10.1016/0006-8993(84)90737-6.
Granule cells from 8-day-old rat cerebella were grown in basal Eagle's medium with 10% fetal calf serum, for 2,5,8 or 12 days in vitro (DIV), in conditions giving a purity greater than 90%. The results obtained can be summarized as follows: (1) Light microscopic autoradiography showed that cultured granule cells and their processes can accumulate the glutamate analog [3H]D-aspartate once they have reached an advanced degree of morphological differentiation (8 and 12 DIV), but, even then, only a limited number of cells was heavily labeled. In contrast, astrocytes were heavily labeled at all stages. (2) Calcium-dependent, high [K+]-induced release, or tetrodotoxin-sensitive, veratridine-induced release of [3H]D-aspartate from granule cell-enriched cultures was detectable only in cultures of 8 or 12 DIV. (3) When subject to 3 consecutive depolarizations, cultured granule cells maintained their ability to release [3H]D-aspartate and endogenous glutamate almost unchanged. (4) Newly synthesized [3H]glutamate was autoradiographically localized in both neurons and astrocytes (the latter, however, were not preferentially labeled as with [3H]D-aspartate), but was specifically released from neuronal structures (perikarya and processes) by depolarizing stimuli.
将8日龄大鼠小脑的颗粒细胞在含有10%胎牛血清的基础伊格尔培养基中培养，在体外培养2、5、8或12天（体外培养天数），培养条件下细胞纯度大于90%。所得结果总结如下：(1)光学显微镜放射自显影显示，培养的颗粒细胞及其突起一旦达到形态分化的高级阶段（体外培养8天和12天），就可以积累谷氨酸类似物[3H]D-天冬氨酸，但即便如此，只有有限数量的细胞被大量标记。相比之下，星形胶质细胞在所有阶段都被大量标记。(2)仅在体外培养8天或12天的培养物中，才能检测到富含颗粒细胞的培养物中[3H]D-天冬氨酸的钙依赖性、高[K+]诱导释放或河豚毒素敏感、藜芦碱诱导释放。(3)当受到连续3次去极化作用时，培养的颗粒细胞释放[3H]D-天冬氨酸和内源性谷氨酸的能力几乎保持不变。(4)新合成的[3H]谷氨酸通过放射自显影定位在神经元和星形胶质细胞中（然而，后者不像[3H]D-天冬氨酸那样被优先标记），但通过去极化刺激从神经元结构（胞体和突起）中特异性释放。
