Pares-Herbuté N, Bonet A, Peraldi S, Pin J P, Gabrion J, Astier H, Tapia-Arancibia L
Unité de Neurobiologie Endocrinologique, C.N.R.S. UA 1197, Université de Montpellier II, France.
Brain Res. 1988 May 1;468(1):89-97. doi: 10.1016/0165-3806(88)90011-9.
We examined the effect of non-neuronal cells on somatostatin release from cultured cerebral cortical cells. Three culture models were used: (1) neuron-enriched cultures obtained from cortex of 17-day-old rat embryos and exposed to 10 microM cytosine arabinoside (Ara C) for 48 h between days 3 and 5 after plating; (2) whole cell cultures obtained by using the same protocol but untreated with Ara C; (3) glial primary cultures obtained from newborn rats. We studied: (i) the cellular composition of the cultures by using two astroglial markers: vimentin and glial fibrillary acidic protein (GFAP); (ii) the spontaneous and forskolin-stimulated somatostatin release. In 8-day-old cultures morphological data revealed that Ara C treatment reduced glial cells to 6%. At 7 and 10 days of culture somatostatin spontaneously released from Ara C-treated cells was higher than that measured from untreated cells. On the 17th day of culture, neuron-enriched cultures contained a lower amount of somatostatin than whole cell cultures. Forskolin elicited a dose-dependent release of somatostatin from whole cell cultures, but had no effect on neuron-enriched cultures. Astroglial released media (ARM) from glial primary cultures exposed to forskolin for 20 min induced somatostatin release from neuron-enriched cultures. HPLC analysis of endogenous amino acids of ARM showed that glutamate, glutamine, glycine and alanine were significantly increased after forskolin stimulation. Our results suggest a functional interaction between glial cells and neurons secreting somatostatin.
我们研究了非神经元细胞对培养的大脑皮质细胞释放生长抑素的影响。使用了三种培养模型:(1)从17日龄大鼠胚胎皮质获得的富含神经元的培养物,在接种后第3天至第5天暴露于10 microM阿糖胞苷(Ara C)48小时;(2)采用相同方案但未用Ara C处理的全细胞培养物;(3)从新生大鼠获得的胶质原代培养物。我们研究了:(i)使用两种星形胶质细胞标志物波形蛋白和胶质纤维酸性蛋白(GFAP)来分析培养物的细胞组成;(ii)生长抑素的自发释放和福斯高林刺激后的释放。在8日龄培养物中,形态学数据显示Ara C处理使胶质细胞减少至6%。在培养第7天和第10天,Ara C处理的细胞自发释放的生长抑素高于未处理细胞所测水平。在培养第17天,富含神经元的培养物中生长抑素的含量低于全细胞培养物。福斯高林能引起全细胞培养物中生长抑素呈剂量依赖性释放,但对富含神经元的培养物无影响。来自经福斯高林处理20分钟的胶质原代培养物的星形胶质细胞释放培养基(ARM)可诱导富含神经元的培养物释放生长抑素。对ARM内源性氨基酸的HPLC分析表明,福斯高林刺激后谷氨酸、谷氨酰胺、甘氨酸和丙氨酸显著增加。我们的结果提示胶质细胞与分泌生长抑素的神经元之间存在功能相互作用。