Type 1 Equilibrative Nucleoside Transporter Regulates Ethanol Drinking Through Accumbal N-Methyl-D-Aspartate Receptor Signaling
Type 1 Equilibrative Nucleoside Transporter Regulates Ethanol Drinking Through Accumbal N-Methyl-D-Aspartate Receptor Signaling
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DOI：
10.1016/j.biopsych.2011.02.013
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发表时间：
2011-06-01
影响因子：
10.6
通讯作者：
Choi, Doo-Sup
中科院分区：
文献类型：
--
作者：
Nam, Hyung Wook;Lee, Moonnoh R.;Choi, Doo-Sup
背景资料：缺乏1型平衡型核苷转运蛋白（ENT 1（-/-））的小鼠与野生型同窝小鼠相比，表现出增加的乙醇偏好行为。ENT 1-/-小鼠的这种表型似乎与丘脑核（NAc）中谷氨酸水平的增加相关。然而，鲜为人知的是，下游的后果增加谷氨酸信号在NAc.Methods：要调查删除ENT 1的意义和谷氨酸信号在NAc的影响，我们采用微透析和iTRAQ蛋白质组学。我们使用Western印迹分析验证了改变的蛋白质。然后，我们研究了抑制N-甲基-D-天冬氨酸（NMDA）谷氨酸受体和蛋白激酶C γ（PKC γ）对野生型小鼠饮酒的药理作用。此外，我们在ENT 1（-/-）背景下使用环磷酸腺苷反应元件-β-半乳糖苷酶小鼠研究了体内环磷酸腺苷反应元件结合活性。我们发现，NMDA谷氨酸受体介导的细胞内PKC γ-神经颗粒蛋白-钙-钙调蛋白依赖性蛋白激酶II型信号传导与降低的环磷酸腺苷反应元件结合活性相关，ENT 1（-/-）小鼠。抑制PKC γ促进野生型小鼠的乙醇饮用水平与ENT 1（-/-）小鼠相似。相比之下，NMDA谷氨酸受体拮抗剂减少了ENT 1（-/-）mice.Conclusions的乙醇饮用量：这些研究结果表明，ENT 1的遗传缺失或药理学抑制调节NMDA谷氨酸受体介导的信号传导在NAc，这提供了一个分子基础，根据ENT 1（-/-）小鼠的乙醇偏好行为。
Background: Mice lacking type 1 equilibrative nucleoside transporter (ENT1(-/-)) exhibit increased ethanol-preferring behavior compared with wild-type littermates. This phenotype of ENT1-/- mice appears to be correlated with increased glutamate levels in the nucleus accumbens (NAc). However, little is known about the downstream consequences of increased glutamate signaling in the NAc.Methods: To investigate the significance of the deletion of ENT1 and its effect on glutamate signaling in the NAc, we employed microdialysis and iTRAQ proteomics. We validated altered proteins using Western blot analysis. We then examined the pharmacological effects of the inhibition of the N-methyl-D-aspartate (NMDA) glutamate receptor and protein kinase C gamma (PKC gamma) on alcohol drinking in wild-type mice. In addition, we investigated in vivo cyclic adenosine monophosphate response element binding activity using cyclic adenosine monophosphate response element-beta-galactosidase mice in an ENT1(-/-) background.Results: We identified that NMDA glutamate receptor-mediated downregulation of intracellular PKC gamma-neurogranin-calcium-calmodulin dependent protein kinase type II signaling is correlated with reduced cyclic adenosine monophosphate response element binding activity in ENT1(-/-) mice. Inhibition of PKC gamma promotes ethanol drinking in wild-type mice to levels similar to those of ENT1(-/-) mice. In contrast, an NMDA glutamate receptor antagonist reduces ethanol drinking of ENT1(-/-) mice.Conclusions: These findings demonstrate that the genetic deletion or pharmacological inhibition of ENT1 regulates NMDA glutamate receptor-mediated signaling in the NAc, which provides a molecular basis that underlies the ethanol-preferring behavior of ENT1(-/-) mice.