Aims: the study aimed to (i) use adeno-associated virus technology to modulate parvalbumin (PV) gene expression, both through overexpression and silencing, within the hippocampus of male mice and (ii) assess the impact of PV on the metabolic pathway of glutamate and γ-aminobutyric acid (GABA). Methods: a status epilepticus (SE) mouse model was established by injecting kainic acid into the hippocampus of transgenic mice. When the seizures of mice reached SE, the mice were killed at that time point and 30 min after the onset of SE. Hippocampal tissues were extracted and the mRNA and protein levels of PV and the 65 kDa (GAD65) and 67 kDa (GAD67) isoforms of glutamate decarboxylase were assessed using real-time quantitative polymerase chain reaction and Western blot, respectively. The concentrations of glutamate and GABA were detected with high-performance liquid chromatography (HPLC), and the intracellular calcium concentration was detected using flow cytometry. Results: we demonstrate that the expression of PV is associated with GAD65 and GAD67 and that PV regulates the levels of GAD65 and GAD67. PV was correlated with calcium concentration and GAD expression. Interestingly, PV overexpression resulted in a reduction in calcium ion concentration, upregulation of GAD65 and GAD67, elevation of GABA concentration, reduction in glutamate concentration, and an extension of seizure latency. Conversely, PV silencing induced the opposite effects. Conclusion: parvalbumin may affect the expression of GAD65 and GAD67 by regulating calcium ion concentration, thereby affecting the metabolic pathways associated with glutamate and GABA. In turn, this contributes to the regulation of seizure activity.

中文翻译：

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小白蛋白通过钙离子浓度调节 GAD 表达，影响 Glu-GABA 平衡，改善 PV-Cre 转基因小鼠 KA 诱导的癫痫持续状态


目的：该研究旨在 (i) 使用腺相关病毒技术通过过度表达和沉默来调节雄性小鼠海马内的小清蛋白 (PV) 基因表达，以及 (ii) 评估 PV 对小清蛋白代谢途径的影响谷氨酸和γ-氨基丁酸（GABA）。方法：通过向转基因小鼠海马注射红藻氨酸建立癫痫持续状态（SE）小鼠模型。当小鼠的癫痫发作达到SE时，在该时间点和SE发作后30分钟处死小鼠。提取海马组织，并分别使用实时定量聚合酶链式反应和蛋白质印迹评估 PV 的 mRNA 和蛋白质水平以及谷氨酸脱羧酶的 65 kDa (GAD65) 和 67 kDa (GAD67) 亚型。采用高效液相色谱法（HPLC）检测谷氨酸和GABA浓度，流式细胞仪检测细胞内钙离子浓度。结果：我们证明 PV 的表达与 GAD65 和 GAD67 相关，并且 PV 调节 GAD65 和 GAD67 的水平。 PV 与钙浓度和 GAD 表达相关。有趣的是，PV过表达导致钙离子浓度降低、GAD65和GAD67上调、GABA浓度升高、谷氨酸浓度降低以及癫痫潜伏期延长。相反，PV 沉默会产生相反的效果。结论：小清蛋白可能通过调节钙离子浓度影响GAD65和GAD67的表达，从而影响谷氨酸和GABA相关的代谢途径。反过来，这有助于调节癫痫活动。