High Ethanol and Acetaldehyde Inhibit Glutamatergic Transmission in the Hippocampus of Aldh2-Knockout and C57BL/6N Mice: an In Vivo and Ex Vivo Analysis.
High Ethanol and Acetaldehyde Inhibit Glutamatergic Transmission in the Hippocampus of Aldh2-Knockout and C57BL/6N Mice: an In Vivo and Ex Vivo Analysis.
复制标题
高乙醇和乙醛抑制 Aldh2 敲除小鼠和 C57BL/6N 小鼠海马中的谷氨酸传输：体内和离体分析。
DOI：
10.1007/s12640-020-00180-6
复制
发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Kinoshita H.
中科院分区：
文献类型：
--
作者：
Jamal M;Ito A;Tanaka N;Miki T;Ameno K;Kinoshita H.
我们的目的是研究乙醇（EtOH）和乙醛（AcH）是否可以影响谷氨酸及其受体GluN 1和GluA 1在Aldh 2基因敲除（Aldh 2-KO）和C57 BL/6 N（野生型（WT））小鼠海马。为此，我们首先检查了局部给予EtOH（100 mM、200 mM和500 mM）和AcH（100 μM、200 μM和500 μM）对自由活动小鼠细胞外谷氨酸水平的影响。将200 mM和500 mM EtOH反向透析到WT和Aldh 2-KO小鼠的海马体中产生细胞外谷氨酸水平的显著降低（p< 0.05）。500 mM EtOH的剂量在Aldh 2-KO小鼠中诱导比在WT小鼠中更大的降低（p< 0.05），表明AcH的作用。类似地，灌注200 μM和500 μM AcH降低了Aldh 2-KO小鼠中的谷氨酸（p< 0.05），但在任何AcH剂量的WT小鼠中均未观察到这种降低。其次，我们测试了乙醇和乙酰胆碱诱导的谷氨酸减少是否与GluN 1和GluA 1表达减少相关，如通过实时PCR和Western印迹所测量的。我们发现WT小鼠在高剂量EtOH（4.0 g/kg）和AcH（200 mg/kg）后GluN 1（p<0.05）和GluA 1（p< 0.05）亚基显著降低。然而，2.0 g/kg剂量的EtOH并未在信使RNA和蛋白质之间产生GluN 1或GluA 1的一致降低。在Aldh 2-KO小鼠中，所有三种剂量的EtOH（1.0 g/kg、2.0 g/kg和4.0 g/kg）和乙酰胆碱（50 mg/kg、100 mg/kg和200 mg/kg）降低GluN 1表达（p< 0.05），而中等至高剂量的EtOH（2.0 g/kg和4.0 g/kg）和AcH（100 mg/kg和200 mg/kg）降低GluA 1表达（p< 0.05）。总之，这些在体内和离体数据表明，乙醇和乙酰胆碱减少细胞外谷氨酸在小鼠海马中的GluN 1和GluA 1亚基伴随减少，但这些效果需要相对较高的浓度，因此，可能解释乙醇中毒的后果。
We aimed to investigate whether ethanol (EtOH) and acetaldehyde (AcH) can affect glutamate and its receptors GluN1 and GluA1 in the hippocampus of Aldh2-knockout (Aldh2-KO) and C57BL/6N (wild-type (WT)) mice. To do this, we first examined the effect of local administration of EtOH (100 mM, 200 mM, and 500 mM) and AcH (100 μM, 200 μM, and 500 μM) on extracellular glutamate levels in freely moving mice. Retrodialysis of 200 mM and 500 mM EtOH into the hippocampus of WT and Aldh2-KO mice produced significant decreases in extracellular glutamate levels (p< 0.05). A dose of 500 mM EtOH induced a greater decrease in Aldh2-KO mice (p< 0.05) than in WT mice, indicating the action of AcH. Similarly, perfusion of 200 μM and 500 μM AcH decreased glutamate in Aldh2-KO mice (p< 0.05), but this decrease was not seen in WT mice at any AcH dose. Second, we tested whether the EtOH- and AcH-induced decrease in glutamate was associated with decreases in GluN1 and GluA1 expression, as measured by real-time PCR and Western blot. We found a significant decrease in GluN1 (p< 0.05) and GluA1 (p< 0.05) subunits after a high dose of EtOH (4.0 g/kg) and AcH (200 mg/kg) in WT mice. However, a 2.0 g/kg dose of EtOH did not produce a consistent decrease in GluN1 or GluA1 between messenger RNA and protein. In Aldh2-KO mice, all three doses of EtOH (1.0 g/kg, 2.0 g/kg, and 4.0 g/kg) and AcH (50 mg/kg, 100 mg/kg, and 200 mg/kg) decreased GluN1 expression (p< 0.05), while moderate-to-high doses of EtOH (2.0 g/kg and 4.0 g/kg) and AcH (100 mg/kg and 200 mg/kg) decreased GluA1 expression (p< 0.05). Together, these in vivo and ex vivo data suggest that EtOH and AcH decrease extracellular glutamate in the hippocampus of mice with a concomitant decrease in GluN1 and GluA1 subunits, but these effects require relatively high concentrations and may, therefore, explain the consequences of EtOH intoxication.