Hacimuftuoglu Ahmet, Tatar Abdulgani, Cetin Damla, Taspinar Numan, Saruhan Fatih, Okkay Ufuk, Turkez Hasan, Unal Deniz, Stephens Robert Louis, Suleyman Halis
Department of Medical Pharmacology, Faculty of Medicine, Atatürk University, Erzurum, Turkey.
Department of Medical Genetics, Faculty of Medicine, Ataturk University, Erzurum, Turkey.
Cytotechnology. 2016 Aug;68(4):1425-33. doi: 10.1007/s10616-015-9902-9. Epub 2015 Oct 6.
The purpose of this study was to clarify the relationship between neuron cells and astrocyte cells in regulating glutamate toxicity on the 10th and 20th day in vitro. A mixed primary culture system from newborn rats that contain cerebral cortex neurons cells was employed to investigate the glutamate toxicity. All cultures were incubated with various glutamate concentrations, then viability tests and histological analyses were performed. The activities of glutamate transporters were determined by using in vitro voltammetry technique. Viable cell number was decreased significantly on the 10th day at 10(-7) M and at 10(-6) M glutamate applications, however, viable cell number was not decreased at 20th day. Astrocyte number was increased nearly six times on the 20th day as compared to the 10th day. The peak point of glutamate reuptake capacity was about 2 × 10(-4) M on the 10th day and 10(-3) M on the 20th day. According to our results, we suggested that astrocyte age was important to maintain neuronal survival against glutamate toxicity. Thus, we revealed activation or a trigger point of glutamate transporters on astrocytes due to time since more glutamate was taken up by astrocytes when glutamate transporters on the astrocyte were triggered with high exogenous glutamate concentrations. In conclusion, the present investigation is the first voltammetric study on the reuptake parameters of glutamate in vitro.
本研究的目的是阐明体外培养第10天和第20天时神经元细胞和星形胶质细胞在调节谷氨酸毒性方面的关系。采用含有大脑皮层神经元细胞的新生大鼠混合原代培养系统来研究谷氨酸毒性。所有培养物均用不同浓度的谷氨酸孵育,然后进行活力测试和组织学分析。采用体外伏安法技术测定谷氨酸转运体的活性。在第10天,当应用10⁻⁷ M和10⁻⁶ M谷氨酸时,活细胞数量显著减少,然而,在第20天活细胞数量并未减少。与第10天相比,第20天星形胶质细胞数量增加了近6倍。第10天谷氨酸重摄取能力的峰值约为2×10⁻⁴ M,第20天为10⁻³ M。根据我们的结果,我们认为星形胶质细胞的年龄对于维持神经元抵抗谷氨酸毒性的存活很重要。因此,我们揭示了由于时间因素,星形胶质细胞上谷氨酸转运体的激活或触发点,因为当星形胶质细胞上的谷氨酸转运体被高浓度外源性谷氨酸触发时,星形胶质细胞会摄取更多的谷氨酸。总之,本研究是首次关于体外谷氨酸重摄取参数的伏安法研究。