Data_Sheet_1_Time dependent changes in protein expression induced by intermittent theta burst stimulation in a cell line.xlsx

BackgroundIntermittent Theta Burst Stimulation (iTBS), a non-invasive brain stimulation technique, is recognized for its ability to modulate cortical neuronal activity. However, its effects over time and the dynamics following stimulation are less well understood. Understanding the temporal dynamics of iTBS effects is essential for optimizing the timing and frequency of stimulation in therapeutic applications.
ObjectiveThis study investigated the temporal changes in protein expression induced by iTBS in Neuro-2a cells.
MethodsWe analyzed protein expression in retinoic acid-differentiated Neuro-2a cells at multiple time points — 0.5, 3, 6, 12, and 24 hours post-iTBS — using Western blot and immunocytochemistry techniques.
ResultsOur findings reveal a significant early increase in neurotransmitter receptor subunits, neurotrophic factors, and cytoskeletal proteins within the first 0.5 hour following iTBS. Notably, proteins such as mGLuR1, NMDAR1, GABBR2, and β-tubulin III showed substantial increase in expression. However, the effects of iTBS on protein expression was not sustained at later timepoints.
ConclusionOur results suggest that iTBS can transiently alter the expression of specific proteins in Neuro-2a cells. Future research should investigate the potential benefits of repeated stimulations within the early time window to refine iTBS interventions, potentially expanding their research and clinical applications.

【背景】间歇性θ爆发刺激（Intermittent Theta Burst Stimulation, iTBS）作为一种非侵入性脑刺激技术，已被证实可调节皮层神经元活动。然而，其随时间推移的效应及刺激后的动态变化尚未得到充分阐明。阐明iTBS效应的时间动态特征，对于优化治疗应用中的刺激时机与频率至关重要。
【目的】本研究旨在探究iTBS诱导Neuro-2a细胞（Neuro-2a）中蛋白质表达的时间变化规律。
【方法】本研究采用蛋白质印迹（Western blot）与免疫细胞化学（immunocytochemistry）技术，在iTBS处理后的0.5、3、6、12及24小时多个时间点，对视黄酸（retinoic acid）诱导分化的Neuro-2a细胞的蛋白质表达水平进行分析。
【结果】研究结果显示，在iTBS处理后的最初0.5小时内，神经递质受体亚基、神经营养因子及细胞骨架蛋白的表达即出现显著上调。其中，mGLuR1、NMDAR1、GABBR2及β-微管蛋白III（β-tubulin III）等蛋白的表达水平升高尤为显著。但在后续时间点，iTBS对蛋白质表达的调控效应并未持续存在。
【结论】本研究结果表明，iTBS可短暂改变Neuro-2a细胞中特定蛋白质的表达水平。未来研究可聚焦于早期时间窗内重复刺激的潜在获益，以优化iTBS干预方案，进而拓展其科研与临床应用场景。