GANGLIOSIDES PREVENT GLUTAMATE AND KAINATE NEUROTOXICITY IN PRIMARY NEURONAL CULTURES OF NEONATAL RAT CEREBELLUM AND CORTEX
GANGLIOSIDES PREVENT GLUTAMATE AND KAINATE NEUROTOXICITY IN PRIMARY NEURONAL CULTURES OF NEONATAL RAT CEREBELLUM AND CORTEX
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DOI：
10.1073/pnas.85.19.7351
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发表时间：
1988-10-01
影响因子：
11.1
通讯作者：
COSTA, E
中科院分区：
文献类型：
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作者：
FAVARON, M;MANEV, H;COSTA, E
使用一个敏感的组织荧光染色方法，允许定量的神经元死亡，我们比较了神经节苷脂（一组天然存在的鞘糖脂），苯环己哌啶（PCP），MK-801（二苯并环庚烯亚胺）谷氨酸和红藻氨酸诱导的神经元死亡的保护作用，在原代培养的皮层和小脑神经元制备的新生大鼠。PCP和MK-801阻断由比LD 50高50倍的谷氨酸剂量（在无Mg 2+的培养基中的LD 50，10 μ M）诱导的神经毒性，但仅部分阻断红藻氨酸神经毒性（在Mg 2+存在下的LD 50，100 μ M）。与此相反，神经节苷脂（GT 1b> GD 1b> GM 1）预处理导致对谷氨酸或红藻氨酸引起的神经毒性的完全和不可逾越的保护。在小脑颗粒细胞的原代培养物中，与PCP和MK-801不同，神经节苷脂未能阻断谷氨酸门控阳离子电流和谷氨酸诱发的（i）肌醇磷脂水解、（ii）c-fos mRNA含量和（iii）c-fos蛋白的核积累的增加。神经节苷脂对谷氨酸神经毒性的保护不需要它们存在于孵育介质中;然而，它与神经元膜中积累的鞘糖脂的量成比例。阻断谷氨酸引起的神经元死亡的神经节苷脂浓度（30-60 μ M）也阻止谷氨酸和红藻氨酸诱导的蛋白激酶C从胞质溶胶易位到神经元膜。
Using a sensitive histofluorescence staining method that allows for a quantitation of neuronal death, we compared the protective effects of gangliosides (a group of naturally occurring glycosphingolipids), phencyclidine (PCP), and MK-801 (dibenzocyclohepteneimine) on glutamate- and kainate-induced neuronal death in primary cultures of cortical and cerebellar neurons prepared from neonatal rats. PCP and MK-801 block neurotoxicity induced by glutamate doses 50 times higher than the LD50 (LD50 in Mg2+-free medium, 10 .mu.M) but only partially block the kainate neurotoxicity (LD50 in presence of Mg2+, 100 .mu.M). In contrast, pretreatment with gangliosides (GT1b > GD1b > GM1) results in complete and insurmountable protection against the neurotoxicity elicited by glutamate or kainate. In primary cultures of cerebellar granule cells gangliosides, unlike PCP and MK-801, fail to block glutamate-gated cationic currents and the glutamate-evoked increase of (i) inositol phospholipid hydrolysis, (ii) c-fos mRNA content, and (iii) nuclear accumulation of c-fos protein. Protection of glutamate neurotoxicity by gangliosides does not require their presence in the incubation medium; however, it is proportional to the amount of glycosphingolipid accumulated in the neuronal membranes. The ganglioside concentration (30-60 .mu.M) that blocks glutamate-elicited neuronal death also prevents glutamate- and kainate-induced protein kinase C translocation from cytosol to neuronal membranes.