An in vitro model for studying the effects of continuous ethanol exposure on N-methyl-D-aspartate receptor function.
An in vitro model for studying the effects of continuous ethanol exposure on N-methyl-D-aspartate receptor function.
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用于研究连续乙醇暴露对 N-甲基-D-天冬氨酸受体功能影响的体外模型。
DOI：
10.1016/j.alcohol.2011.08.003
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发表时间：
2012-02
期刊：
影响因子：
--
通讯作者：
Popp RL
中科院分区：
文献类型：
--
作者：
Nath V;Reneau JC;Dertien JS;Agrawal RG;Guerra I;Bhakta Y;Busari K;Neumann MK;Bergeson SE;Popp RL
长期乙醇暴露对神经胶质和神经元功能都有有害影响。我们评估了连续乙醇暴露（CEE）后大鼠小脑颗粒细胞（CGCs）和星形胶质细胞共培养物中星形胶质细胞和神经元活力的变化，以及N-甲基-d-天冬氨酸受体（NMDAR）功能的变化。每24小时用100 mM EtOH处理细胞一次，持续四天，导致平均乙醇浓度为57.3 ± 2.1 mM。在最后一次乙醇处理后四天对对照和乙醇处理后的细胞进行比较。CEE没有改变神经胶质细胞的活力，如在乙醇处理后的任何一天星形胶质细胞形态，肌动蛋白解聚或星形胶质细胞内线粒体分布的破坏的情况下的变化所示。相差显微镜和台盼蓝拒染法显示，CEE后的CGCs是健康和存活的。全细胞膜片钳实验表明，NMDA诱导的电流（INMDA）改变CEE处理。类似于先前的结果，在戒断期从慢性乙醇暴露，INMDA从CEE处理的细胞显着大于INMDA从NMDARs在控制CGCs，但返回到控制值的第四天后CEE。然而，在最后一次乙醇给药后和乙醇浓度保持较高的时间内，INMDA显著小于对照值。在表达NR 2A或NR 2B亚基的CGCs中观察到相同的结果。总之，神经元和星形胶质细胞在暴露于CEE后保持健康，在细胞水平上没有神经毒性的迹象，并且NMDAR功能的调节与先前实验的发现一致。因此，我们得出的结论是，神经胶质-神经元共培养物中的连续乙醇暴露范式很容易适用于乙醇影响的长期体外研究，包括神经胶质-神经元相互作用和研究乙醇戒断诱导的神经毒性的能力。
Long-term ethanol exposure has deleterious effects on both glial and neuronal function. We assessed alterations in both astrocytic and neuronal viability, as well as alterations in N-methyl-d-aspartate receptor (NMDAR) function, in co-cultures of rat cerebellar granule cells (CGCs) and astrocytes after continuous ethanol exposure (CEE). Treatment of cells with 100 mM EtOH once every 24 h for four days resulted in a mean ethanol concentration of 57.3 ± 2.1 mM. Comparisons between control and post-ethanol treated cells were made four days after the last ethanol treatment. CEE did not alter glial cell viability, as indicated by the absence of either changes in astrocytic morphology, actin depolymerization, or disruption of astrocytic intracellular mitochondrial distribution at any day post-ethanol treatment. The CGCs were healthy and viable after CEE, as indicated by phase contrast microscopy and the trypan blue exclusion method. Whole-cell patch-clamp experiments indicated that NMDA-induced currents (INMDA) were altered by CEE treatment. Similar to previous results obtained during the withdrawal phase from chronic ethanol exposure, INMDA from CEE-treated cells were significantly larger than INMDA from NMDARs in control CGCs, but returned to control values by the fourth day post-CEE. However, after the last ethanol dosing and during a time when ethanol concentrations remained high, INMDA were significantly smaller than control values. Identical results were observed in CGCs expressing the NR2A or NR2B subunit. In summary, both neurons and astrocytes remained healthy following exposure to CEE with no signs of neurotoxicity at the cellular level, and modulation of NMDAR function are consistent with findings from prior experiments. Thus, we conclude that the continuous ethanol exposure paradigm in glial-neuronal co-cultures readily lends itself to long-term in vitro studies of ethanol effects that include glial-neuronal interactions and the ability to study ethanol withdrawal-induced neurotoxicity.
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DOI：
10.1038/npp.2009.69
发表时间：
2009-10
期刊：
Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology
影响因子：
--
作者：
通讯作者：
--
影响因子：
4.2
作者：
Guasch, RM;Tomas, M;Guerri, C
通讯作者：
Guerri, C
影响因子：
2.1
作者：
Hatton, GI
通讯作者：
Hatton, GI
影响因子：
4.8
作者：
Follesa, P;Ticku, MK
通讯作者：
Ticku, MK
影响因子：
56.9
作者：
LOVINGER, DM;WHITE, G;WEIGHT, FF
通讯作者：
WEIGHT, FF