Chloride channels with ClC-1-like properties differentially regulate the excitability of dopamine receptor D1- and D2-expressing striatal medium spiny neurons.
Chloride channels with ClC-1-like properties differentially regulate the excitability of dopamine receptor D1- and D2-expressing striatal medium spiny neurons.
复制标题
具有 ClC-1 样特性的氯离子通道差异调节表达多巴胺受体 D1 和 D2 的纹状体中型多棘神经元的兴奋性。
DOI：
10.1152/ajpcell.00397.2021
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Hauser,KurtF
中科院分区：
文献类型：
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作者：
Yarotskyy,Viktor;Lark,AriannaRS;Nass,SaraR;Hahn,YunK;Marone,MichaelG;McQuiston,ARory;Knapp,PamelaE;Hauser,KurtF
动态氯 (Cl−) 调节对于突触抑制至关重要。在成熟神经元中，Cl−流入和挤出主要分别由配体门控阴离子通道（GABAA 和甘氨酸受体）和氯化钾协同转运蛋白 K+-Cl−协同转运蛋白 2 (KCC2) 控制。在这里，据我们所知，我们首次报告纹状体神经元中存在一种新的 Cl−流入源，其特性类似于氯电压门控通道 1 (ClC-1)。使用全细胞膜片钳记录，我们检测到一种向外整流的电压依赖性电流，该电流对于大阴离子甲磺酸盐（MsO−）是不可渗透的。阴离子电流对 ClC-1 抑制剂 9-蒽甲酸 (9-AC) 和非特异性阻断剂根皮素敏感。 MsO−、9-AC 和根皮素抑制阴离子电流的平均分数没有显着差异，表明阴离子电流是由活跃的 ClC-1 样通道引起的。此外，我们发现 Cl− 电流对跨膜蛋白 16A (TMEM16A;Ano1) 抑制剂 Ani9 不敏感，并且即使在非常高的细胞内 Ca2+ 浓度 (2 mM) 下，向外的 Cl− 整流也得以保留，表明 TMEM16B (Ano2) 对总电流没有贡献。 Western印迹和免疫组织化学分析证实纹状体中存在ClC-1通道，主要定位于纹状体神经元的体细胞。最后，我们发现 9-AC 降低了脑切片中分别表达 1 型多巴胺 (D1) 和 2 型 (D2) 受体的中型多巴胺神经元 (MSN) 的动作电位放电频率并增加了兴奋性。我们得出结论，ClC-1 样通道优先位于 MSN 体细胞，具有功能性，并且可以调节神经元兴奋性。
Dynamic chloride (Cl−) regulation is critical for synaptic inhibition. In mature neurons, Cl−influx and extrusion are primarily controlled by ligand-gated anion channels (GABAAand glycine receptors) and the potassium chloride cotransporter K+-Cl−cotransporter 2 (KCC2), respectively. Here, we report for the first time, to our knowledge, a presence of a new source of Cl−influx in striatal neurons with properties similar to chloride voltage-gated channel 1 (ClC-1). Using whole cell patch-clamp recordings, we detected an outwardly rectifying voltage-dependent current that was impermeable to the large anion methanesulfonate (MsO−). The anionic current was sensitive to the ClC-1 inhibitor 9-anthracenecarboxylic acid (9-AC) and the nonspecific blocker phloretin. The mean fractions of anionic current inhibition by MsO−, 9-AC, and phloretin were not significantly different, indicating that anionic current was caused by active ClC-1-like channels. In addition, we found that Cl−current was not sensitive to the transmembrane protein 16A (TMEM16A;Ano1) inhibitor Ani9 and that the outward Cl−rectification was preserved even at a very high intracellular Ca2+concentration (2 mM), indicating that TMEM16B (Ano2) did not contribute to the total current. Western blotting and immunohistochemical analyses confirmed the presence of ClC-1 channels in the striatum mainly localized to the somata of striatal neurons. Finally, we found that 9-AC decreased action potential firing frequencies and increased excitability in medium spiny neurons (MSNs) expressing dopamine type 1 (D1) and type 2 (D2) receptors in the brain slices, respectively. We conclude that ClC-1-like channels are preferentially located at the somata of MSNs, are functional, and can modulate neuronal excitability.