Barr Jeffrey L, Deliu Elena, Brailoiu G Cristina, Zhao Pingwei, Yan Guang, Abood Mary E, Unterwald Ellen M, Brailoiu Eugen
Center for Substance Abuse Research, Temple University School of Medicine, Philadelphia, PA 19140, USA; Department of Pharmacology, Temple University School of Medicine, Philadelphia, PA 19140, USA.
Center for Substance Abuse Research, Temple University School of Medicine, Philadelphia, PA 19140, USA.
Cell Calcium. 2015 Aug;58(2):196-207. doi: 10.1016/j.ceca.2015.05.001. Epub 2015 May 27.
Cocaine promotes addictive behavior primarily by blocking the dopamine transporter, thus increasing dopamine transmission in the nucleus accumbens (nAcc); however, additional mechanisms are continually emerging. Sigma-1 receptors (σ1Rs) are known targets for cocaine, yet the mechanisms underlying σ1R-mediated effects of cocaine are incompletely understood. The present study examined direct effects of cocaine on dissociated nAcc neurons expressing phosphatidylinositol-linked D1 receptors. Endoplasmic reticulum-located σ1Rs and inositol 1,4,5-trisphosphate (IP3) receptors (IP3Rs) were targeted using intracellular microinjection. IP3 microinjection robustly elevated intracellular Ca(2+) concentration, [Ca(2+)]i. While cocaine alone was devoid of an effect, the IP3-induced response was σ1R-dependently enhanced by cocaine co-injection. Likewise, cocaine augmented the [Ca(2+)]i increase elicited by extracellularly applying an IP3-generating molecule (ATP), via σ1Rs. The cocaine-induced enhancement of the IP3/ATP-mediated Ca(2+) elevation occurred at pharmacologically relevant concentrations and was mediated by transient receptor potential canonical channels (TRPC). IP3 microinjection elicited a slight, transient depolarization, further converted to a greatly enhanced, prolonged response, by cocaine co-injection. The cocaine-triggered augmentation was σ1R-dependent, TRPC-mediated and contingent on [Ca(2+)]i elevation. ATP-induced depolarization was similarly enhanced by cocaine. Thus, we identify a novel mechanism by which cocaine promotes activation of D1-expressing nAcc neurons: enhancement of IP3R-mediated responses via σ1R activation at the endoplasmic reticulum, resulting in augmented Ca(2+) release and amplified depolarization due to subsequent stimulation of TRPC. In vivo, intra-accumbal blockade of σ1R or TRPC significantly diminished cocaine-induced hyperlocomotion and locomotor sensitization, endorsing a physio-pathological significance of the pathway identified in vitro.
可卡因主要通过阻断多巴胺转运体来促进成瘾行为,从而增加伏隔核(nAcc)中的多巴胺传递;然而,其他机制也在不断涌现。西格玛-1受体(σ1Rs)是可卡因已知的作用靶点,但可卡因通过σ1R介导的效应的潜在机制尚未完全明确。本研究检测了可卡因对表达磷脂酰肌醇连接的D1受体的离体伏隔核神经元的直接作用。通过细胞内微注射靶向内质网定位的σ1Rs和肌醇1,4,5-三磷酸(IP3)受体(IP3Rs)。微注射IP3可显著提高细胞内钙离子浓度[Ca(2+)]i。单独使用可卡因没有效果,但联合注射可卡因可通过σ1R依赖性增强IP3诱导的反应。同样,可卡因通过σ1Rs增强了细胞外应用产生IP3的分子(ATP)所引起的[Ca(2+)]i升高。可卡因诱导的IP3/ATP介导的钙离子升高增强发生在药理学相关浓度,并且由瞬时受体电位经典通道(TRPC)介导。微注射IP3引起轻微的、短暂的去极化,联合注射可卡因进一步将其转化为大大增强的、延长的反应。可卡因引发的增强作用是σ1R依赖性的、TRPC介导的,并且取决于[Ca(2+)]i升高。ATP诱导的去极化同样被可卡因增强。因此,我们确定了一种可卡因促进表达D1的伏隔核神经元激活的新机制:通过在内质网激活σ1R增强IP3R介导的反应,导致钙离子释放增加,并由于随后对TRPC的刺激而使去极化放大。在体内,伏隔核内阻断σ1R或TRPC可显著减少可卡因诱导的过度活动和运动敏化,证实了体外确定的该途径的生理病理意义。