Mechanisms involved in the Development Behavioural supersensitivity

短期目标和长期目标：该项目的短期目标是：(1)深入了解多巴胺D2受体（D2R）拮抗后行为超敏感发展的分子机制；(2)研究我们实验室设计的新型肽A的保护机制。长期目标是：(3)利用活体大鼠的脑成像研究氧化应激（OS）诱导的多巴胺转运体和受体的变化；(4)通过开发新型微凝胶靶向递送肽A到大脑来增强神经保护。****目标与途径****短期5年；为了研究新型肽A对氟哌啶醇诱导的OS的神经保护作用，我们将不同组的大鼠分别给予氟哌啶醇、氟哌啶醇+肽A和无活性肽B治疗28天。行为超敏感将通过记录刻板行为、真空咀嚼运动（VCMs）和运动活动来监测。2. 为了确定肽A是否能阻止AIF在纹状顶神经元、纹状间管神经元和星形胶质细胞中的易位，将各组大鼠分别给予氟哌啶醇、氟哌啶醇+肽A和载药液治疗28天。在治疗结束时，大脑区域将被解剖，并用针对神经元和星形胶质细胞的特异性抗体进行标记。3. 为了确定是否存在谷氨酸释放增加和GABA减少的情况，将按照上述目的2对大鼠进行处理。用微透析法测定活体大鼠的谷氨酸和氨基丁酸。4. 为了研究多肽A在细胞模型中的分子机制，将使用d2r转染的人神经母细胞瘤SH SY5Y细胞检测环境毒素、百草草和氟哌啶醇处理后的自由基形成、MEF2和Nrf2表达以及细胞活力。**** 5年以上的长期目标：利用脑成像技术确定肽A的转运是否能阻止氧化应激过程中多巴胺转运体和D2Rs的下调。2. 利用我们的专利技术（专利号62/362,105,2016）开发用于靶向递送肽A到大脑的新型微凝胶。****新颖性，重要性和影响：****该计划使用从细胞模型到动物试验的技术来推进我们对氧化应激和细胞死亡的认识。由于某些化合物和环境毒素引起的细胞死亡机制尚不清楚，因此这项研究至关重要。此外，对神经保护的新认识将有助于促进更有效化合物的开发。结合用于增强靶向脑输送的新型微凝胶的生产，该项目将开发的可专利技术为加拿大进一步增加其对新型生物和工程知识和技术的贡献提供了令人兴奋的机会

Short Term Objectives and Long Term Goals: The short term objectives of this program are to (1) gain an in-depth understanding of the molecular mechanisms involved in the development of behavioural supersensitivity following dopamine D2 receptor (D2R) antagonism and (2) to investigate the protective mechanisms of novel Peptide A designed in our lab. The long-term goals are to (3) investigate the changes in dopamine transporters and receptors induced by oxidative stress (OS) using brain imaging in live rats and (4) to enhance neuroprotection by developing novel microgels for targeted delivery of Peptide A to the brain.****Aims & Approaches****Short term 5 years: 1. To investigate the effects of novel Peptide A for neuroprotection against haloperidol-induced OS, various groups of rats will be treated with haloperidol, haloperidol + Peptide A, and inactive Peptide B for 28 days. Behavioural supersensitivity will be monitored by recording stereotyped behavior, vacuous chewing movements (VCMs) and locomotor activity. 2. To establish whether Peptide A prevents AIF translocation in striatopallidal neurons, striatoentopenduncular neurons, and astroglial cells, groups of rats will be treated with haloperidol, haloperidol + Peptide A, and vehicle solution for 28 days. At the end of treatment, brain regions will be dissected and labeled with specific antibodies for neurons and astroglial cells. 3. To establish whether there is an increase in glutamate release and decrease in GABA, rats will be treated as described above in Aim 2. Glutamate and GABA will be measured in live rats by microdialysis. 4. To investigate molecular mechanisms of Peptide A in cellular models, D2R-transfected human neuroblastoma SH SY5Y cells will be used to measure free radical formation, MEF2 and Nrf2 expression, and cell viability upon treatment with environmental toxin, paraquat, and haloperidol.****Long-term goals, beyond 5 years: 1. To determine whether intransal delivery of Peptide A prevents downregulation of dopamine transporters and D2Rs during oxidative stress using brain imaging. 2. To develop novel microgels for targeted delivery of Peptide A to the brain using our patented technology (Patent # 62/362,105,2016).****Novelty, Significance, and Impact:****This program uses technology ranging from cellular models to animal testing to advance our knowledge of oxidative stress and cell death. This research is essential as the mechanisms involved in cell death caused by certain compounds and environmental toxins are not known. Additionally, new insights into neuroprotection will help advance the development of more effective compounds. Combined with the production of novel microgels for enhanced targeted brain delivery, the patentable technology that will be developed in this project represents exciting opportunities for Canada to further increase its contribution to novel biological and engineering knowledge and techniques.***