Background

Over the course of chronic drug use, brain transcriptional neuroadaptation is thought to contribute to a change in drug use behavior over time. The function of the transcription factor CREB (cAMP response element binding protein) within the nucleus accumbens (NAc) has been well documented in opposing the rewarding properties of many classes of drugs, yet the gene targets through which CREB causally manifests these lasting neuroadaptations remain unknown. Here, we identify zinc finger protein 189 (Zfp189) as a CREB target gene that is transcriptionally responsive to acute and chronic cocaine use within the NAc of mice.

Methods

To investigate the role of the CREB-Zfp189 interaction in cocaine use, we virally delivered modified clustered regularly interspaced short palindromic repeats (CRISPR)/dCas9 constructs capable of selectively localizing CREB to the Zfp189 gene promoter in the NAc of mice.

Results

We observed that CREB binding to the Zfp189 promoter increased Zfp189 expression and diminished the reinforcing responses to cocaine. Furthermore, we showed that NAc Zfp189 expression increased within D1 medium spiny neurons in response to acute cocaine but increased in both D1- and D2-expressing medium spiny neurons in response to chronic cocaine. CREB-mediated induction of Zfp189 potentiated electrophysiological activity of D1- and D2-expressing medium spiny neurons, recapitulating the known effect of CREB on these neurons. Finally, targeting CREB to the Zfp189 promoter within NAc Drd2-expressing neurons, but not Drd1-expressing neurons, was sufficient to diminish cocaine-conditioned behaviors.

Conclusions

Together, these findings point to the CREB-Zfp189 interaction within the NAc Drd2+ neurons as a molecular signature of chronic cocaine use that is causal in counteracting the reinforcing effects of cocaine.

中文翻译：

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CREB ​​与中型多棘神经元亚型内 Zfp189 启动子的结合可差异调节可卡因使用过程中的行为和生理适应

 背景

在长期吸毒过程中，大脑转录神经适应被认为有助于随着时间的推移改变吸毒行为。伏隔核 (NAc) 内的转录因子 CREB（cAMP 反应元件结合蛋白）的功能已被充分证明可以对抗许多类别药物的有益特性，但 CREB ​​因果性地表现出这些持久神经适应的基因靶标仍然未知。在这里，我们将锌指蛋白 189 ( Zfp189 ) 确定为 CREB ​​靶基因，该基因对小鼠 NAc 内的急性和慢性可卡因使用有转录反应。

 方法

为了研究 CREB- Zfp189相互作用在可卡因使用中的作用，我们通过病毒传递修饰的成簇规则间隔短回文重复序列 (CRISPR)/dCas9 构建体，能够选择性地将 CREB ​​定位到小鼠 NAc 中的Zfp189基因启动子。

 结果

我们观察到 CREB ​​与Zfp189启动子的结合增加了Zfp189 的表达并减弱了对可卡因的增强反应。此外，我们发现NAc Zfp189表达在D1中型多棘神经元中对急性可卡因的反应增加，但在D1和D2表达的中型多棘神经元中对慢性可卡因的反应增加。 CREB ​​介导的Zfp189诱导增强了表达 D1 和 D2 的中型多棘神经元的电生理活性，概括了 CREB ​​对这些神经元的已知作用。最后，将 CREB ​​靶向表达 NAc Drd2的神经元内的Zfp189启动子，而不是表达Drd1的神经元，足以减少可卡因条件反射行为。

 结论

总之，这些发现表明 NAc Drd2+神经元内的 CREB- Zfp189相互作用是长期使用可卡因的分子特征，是抵消可卡因增强作用的原因。