Gene Delivery and Addiction

重组腺相关病毒(RAAV)载体通常用于中枢神经系统的基因传递，并且能够转导神经元。使用表达胶质源性神经营养因子(GDNF)的AAV载体，我们能够与我们的内科合作者Yavin Shaham博士一起参与一项研究。我们发现，在可卡因戒断后的第一天注射含有大鼠GDNF基因的腺相关病毒(AAV)病毒载体会增加戒断第11天和第31天的线索诱导的可卡因寻找；这种影响在对照病毒中没有观察到。这项完整的研究考察了GDNF在培养可卡因欲望中的作用。这项取消标题的研究目前正在审查中，准备发表。 我们部门多年来一直在研究的蛋白质之一是骨形态发生蛋白7，特别是它的神经营养/神经再生特性。我们先前证明，外源性应用骨形态发生蛋白7(BMP7)可以减少帕金森病啮齿动物模型中6-羟基多巴胺介导的神经变性。我们在过去一年发表的一项研究表明，在黑质纹状体通路的多巴胺能神经元中表达截短形式的BMP受体II的转基因小鼠更容易受到甲基苯丙胺的攻击，这一点通过黑质的TUNEL标记确定。我们的数据表明，BMP信号的缺陷增加了对高剂量MA引起的侮辱的易感性。在第二项研究中，我们发现甲基苯丙胺抑制了小鼠黑质中BMP7的表达。仅携带BMP7一个等位基因的小鼠更容易受到甲基苯丙胺毒性的影响，这是通过黑质网状体内酪氨酸羟化酶(TH)免疫染色和运动活动的变化来衡量的。通过小鼠侧脑室外源性输送BMP7蛋白减少了甲基苯丙胺的毒性，通过运动活动和TH免疫染色来测量。总之，我们的数据表明，BMP7和BMP受体信号对黑质纹状体系统中的甲基苯丙胺毒性具有神经保护作用。 我们最近建立了一个表达人类MU阿片受体(HUM)的AAV载体，并已开始评估HUM在小鼠甲基苯丙胺敏化中的作用。我们的初步发现表明，AAV载体在特定大脑区域的幽默表达改变了甲基苯丙胺的敏化。这项工作正在进行中。 构建了表达谷氨酸转运体(GLT-1)的AAV载体，以调节细胞外谷氨酸水平。我们已经开始实验检测AAV过量表达GLT-1来减少谷氨酸的兴奋性毒性的能力。我们还开始研究GLT-1在特定脑区的过度表达，以寻找对甲基苯丙胺敏感的变化。一旦完全特征化，AAV-GLT1载体将被用于药物敏化、渴望和奖励的其他模型。

Recombinant adeno-associated viral (rAAV) vectors are frequently used for gene delivery to the central nervous system and are capable of transducing neurons. Using an AAV vector expressing glial derived neurotrophic factor (GDNF), we were able to contribute to a study with our intramural collaborator, Dr. Yavin Shaham. We show that injections of an adeno-associated virus (AAV) viral vector containing rat GDNF cDNA on the first day after cocaine withdrawal increased cue-induced cocaine-seeking on withdrawal days 11 and 31; this effect was not observed using a control virus. The complete study examines the role of GDNF in the incubation of cocaine craving. The detialed study is currently under review for publication. One of the proteins that our section has been studying for many years is bone morphogenetic protein 7, specifically, its neurotrophic/neuroregenerative properties. We previously demonstrated that exogenous application of bone morphogenetic protein 7 (BMP7) reduced 6-hydroxydopamine-mediated neurodegeneration in a rodent model of Parkinson's disease. We published a study this past year showing that transgenic mice expressing a truncated form of the BMP receptor II in dopaminergic neurons of the nigrostriatal pathway were more vulnerable to methamphetamine challenge as determined by TUNEL labeling in the substantia nigra. Our data suggest that a deficiency in BMP signaling increases vulnerability to insults induced by high doses of MA. In a second study, we show that methamphetamine suppressed BMP7 expression in the substantia nigra of mice. Mice that carry only one allele of BMP7 were more vulnerable to methamphetamine toxicity as measured by tyrosine hydroxylase (TH) immunostaining in the nigra reticulate and changes in locomotor activity. Exogenous delivery of BMP7 protein via the lateral ventricle of mouse brain reduced methamphetamine toxicity as measured by locomotor activity and TH immunostaining. Collectively, our data indicate that BMP7 and BMP receptor signaling has neuroprotective effects against methamphetamine toxicity in the nigrostriatal system. We recently generated an AAV vector expressing the human mu opioid receptor (huMOR) and have begun evaluating the role of huMOR in methamphetamine sensitization in mice. Our preliminary findings show that huMOR expression by an AAV vector in specific brain regions alters methamphetamine sensitization. This work is ongoing. An AAV vector expressing the glutamate transporter (GLT-1) was created to modulated the levels of extracellular glutamate. We have begun experiments examining the ability of excess GLT-1 overexpression by AAV to reduce excitoxicity by glutamate. We have also begun examining the GLT-1 overexpression in specfic brain regions for alterations to methamphetamine sensitization. Once fully characterized, the AAV-GLT1 vector will be used in other models of drug sensitization, craving and reward.