细胞培养环境对BV-2小胶质细胞转录因子MafB的调控

Matters Pub Date : 2021-01-01 Epub Date: 2021-01-29
Patrick Miller-Rhodes, Harris A Gelbard
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引用次数: 0

摘要

当小胶质细胞从中枢神经系统(CNS)转移到细胞培养环境时,会经历剧烈的分子和功能变化。研究人员将这些发现很大程度上归因于中枢神经系统特异性微环境线索的缺失,这些微环境线索决定了由主调控转录因子(如V-maf肌肉腱神经纤维肉瘤癌基因同源物B (MafB))指定的基因调控网络。mab通过激活或抑制对这些过程至关重要的靶基因来调节巨噬细胞的分化和激活。在这里,我们发现BV-2小胶质细胞系的基础mab水平取决于细胞培养环境中脂质的可用性。脂质消耗,无论是通过血清剥夺或使用脂质消耗血清,降低BV-2细胞中的MafB蛋白水平。通过实时成像,我们还观察到凋亡的BV-2细胞碎片被邻近的BV-2细胞吞噬,强调了细胞培养环境中脂质的另一个潜在来源。这一观察结果得到了实验的支持,实验显示,用各种吞噬抑制剂(细胞松弛素D、膜联蛋白V)培养的BV-2细胞中,mab蛋白水平降低,血清剥夺后BV-2细胞的吞噬活性降低。总之,我们的数据表明,血清暴露通过直接和间接的机制调节BV-2细胞中的转录因子MafB。
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The Cell Culture Environment Regulates the Transcription Factor MafB in BV-2 Microglia.

Microglia experience dramatic molecular and functional changes when transferred from the central nervous system (CNS) to a cell culture environment. Investigators largely attribute these findings to the loss of CNS-specific microenvironmental cues that dictate the gene-regulatory networks specified by master regulator transcription factors such as V-maf musculoaponeurotic fibrosarcoma oncogene homolog B (MafB). MafB regulates macrophage differentiation and activation by activating or repressing target genes critical to these processes. Here, we show that basal MafB levels in the BV-2 microglial cell line depend on the availability of lipids in the cell culture environment. Depletion of lipids, either by serum deprivation or the use of lipid-depleted serum, reduced MafB protein levels in BV-2 cells. Using live imaging, we also observed the engulfment of apoptotic BV-2 cell debris by neighboring BV-2 cells, highlighting an additional potential source of lipids in the cell culture environment. This observation was supported by experiments showing reduced MafB protein levels in BV-2 cells cultured with various phagocytosis inhibitors (cytochalasin D, annexin V) and reduced BV-2 cell phagocytic activity with serum deprivation. In aggregate, our data suggest that serum exposure regulates the transcription factor MafB in BV-2 cells through direct and indirect mechanisms.

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