Uptake of Biotinylated Spermine in Astrocytes: Effect of Cx43 siRNA, HIV-Tat Protein and Polyamine Transport Inhibitor on Polyamine Uptake.
Uptake of Biotinylated Spermine in Astrocytes: Effect of Cx43 siRNA, HIV-Tat Protein and Polyamine Transport Inhibitor on Polyamine Uptake.
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DOI：
10.3390/biom11081187
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发表时间：
2021-08-11
期刊：
影响因子：
5.5
通讯作者：
Skatchkov SN
中科院分区：
文献类型：
--
作者：
Malpica-Nieves CJ;Rivera Y;Rivera-Aponte DE;Phanstiel O;Veh RW;Eaton MJ;Skatchkov SN
多胺（PAs）是含有多个氨基的多阳离子生物分子。hiv相关神经认知障碍（HAND）患者的大脑和脑脊液（CSF）中存在高浓度的多胺n -乙酰化精胺，并且星形胶质细胞释放的PA增加。这些影响是由于暴露于HIV-Tat。在健康成人大脑中，PAs在星形胶质细胞中积累而不合成，这表明PAs必须进入星形胶质细胞才能被n -乙酰化并释放。因此，我们测试了Cx43半通道（Cx43- hc）在对照和hiv - tat处理的星形胶质细胞中是否是PA通量的途径。我们使用生物素化精胺（b-SPM）检测多胺摄取。我们发现对照星形胶质细胞和siRNA-Cx43处理的星形胶质细胞摄取b-SPM，同样表明PA摄取是通过转运体/通道而不是cx43 - hc。令人惊讶的是，用HIV-Tat和siRNA-Cx43预处理的星形胶质细胞显示b-SPM的积累增加。使用一种新型多胺运输抑制剂（PTI），三聚体44NMe，我们阻断了b-SPM的摄取，表明PA的摄取是通过PTI敏感的运输机制，如有机阳离子转运体。我们的数据表明，在正常的细胞外钙浓度条件下，Cx43 hc不是b-SPM摄取的主要途径，但可能参与病毒感染期间pa向细胞外空间的释放。
Polyamines (PAs) are polycationic biomolecules containing multiple amino groups. Patients with HIV-associated neurocognitive disorder (HAND) have high concentrations of the polyamine N-acetylated spermine in their brain and cerebral spinal fluid (CSF) and have increased PA release from astrocytes. These effects are due to the exposure to HIV-Tat. In healthy adult brain, PAs are accumulated but not synthesized in astrocytes, suggesting that PAs must enter astrocytes to be N-acetylated and released. Therefore, we tested if Cx43 hemichannels (Cx43-HCs) are pathways for PA flux in control and HIV-Tat-treated astrocytes. We used biotinylated spermine (b-SPM) to examine polyamine uptake. We found that control astrocytes and those treated with siRNA-Cx43 took up b-SPM, similarly suggesting that PA uptake is via a transporter/channel other than Cx43-HCs. Surprisingly, astrocytes pretreated with both HIV-Tat and siRNA-Cx43 showed increased accumulation of b-SPM. Using a novel polyamine transport inhibitor (PTI), trimer 44NMe, we blocked b-SPM uptake, showing that PA uptake is via a PTI-sensitive transport mechanism such as organic cation transporter. Our data suggest that Cx43 HCs are not a major pathway for b-SPM uptake in the condition of normal extracellular calcium concentration but may be involved in the release of PAs to the extracellular space during viral infection.
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