Fritz J D, Utz A L, Hale N M, Wu L, Powers A C, Verdoorn T A, Robertson D
Department of Pharmacology, Vanderbilt University, Nashville, Tennessee 37232, USA.
J Neurochem. 1997 Jan;68(1):204-12. doi: 10.1046/j.1471-4159.1997.68010204.x.
The human glutamic acid decarboxylase (GAD) gene was transferred into rat cerebellar granule neurons. Following adenoviral-mediated gene transfer, nearly 100% of the neurons had transgene expression that persisted for the duration of their survival in culture. GABA levels were elevated both in the growth media and in lysates of GAD-modified granule neurons. In GAD-modified neurons, extracellular GABA levels steadily increased with time, whereas intracellular GABA levels peaked 10 days after gene transfer. GAD-modified neurons released both glutamate and GABA into the surrounding media before and after potassium-induced stimulation, but only the release of glutamate was sensitive to potassium stimulation. These data suggest that glutamatergic neurons, which initially contained no detectable GABA, can be genetically modified to release GABA constitutively.
将人类谷氨酸脱羧酶（GAD）基因转入大鼠小脑颗粒神经元。经腺病毒介导的基因转移后，近100%的神经元有转基因表达，且在其培养存活期间持续存在。在生长培养基和GAD修饰的颗粒神经元裂解物中，GABA水平均升高。在GAD修饰的神经元中，细胞外GABA水平随时间稳步增加，而细胞内GABA水平在基因转移后10天达到峰值。在钾离子诱导刺激前后，GAD修饰的神经元将谷氨酸和GABA释放到周围培养基中，但只有谷氨酸的释放对钾离子刺激敏感。这些数据表明最初不含可检测到的GABA的谷氨酸能神经元可通过基因改造组成性释放GABA。
