Mechanisms involved in the Development Behavioural supersensitivity

短期目标和长期目标：该计划的短期目标是：(1)深入了解多巴胺D2受体(D2R)拮抗后行为超敏反应发生的分子机制；(2)研究本实验室设计的新型多肽A的保护机制。长期目标是(3)利用活体大鼠的脑成像技术研究氧化应激(OS)诱导的多巴胺转运体和受体的变化，以及(4)通过开发新型微凝胶将多肽A定向输送到大脑来增强神经保护。 目标和方法 短期5年：1.为探讨新型多肽A对氟哌啶醇诱导的OS的神经保护作用，对不同剂量的大鼠分别给予氟哌啶醇、氟哌啶醇+多肽A和灭活多肽B治疗28天。将通过记录刻板印象的行为、空腹咀嚼运动(VCM)和运动活动来监测行为超敏感。2.为确定多肽A是否能阻止AIF在纹状体苍白质神经元、纹状体丘脑神经元和星形胶质细胞中的易位，大鼠分别给予氟哌啶醇、氟哌啶醇+多肽A和溶媒治疗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定抗体进行标记。3.为了确定是否存在谷氨酸释放增加和GABA减少，将在目标2中对大鼠进行上述处理。4.以D2R基因转染人神经母细胞瘤SH SY5Y细胞为研究对象，检测环境毒素、百草枯、氟哌啶醇对细胞内自由基生成、MEF2、NRF2表达的影响，探讨多肽A在细胞模型中的作用机制。 超过5年的长期目标：1.通过脑成像确定多肽A经腹给药是否能阻止氧化应激期间多巴胺转运体和D2Rs的下调。2.利用我们的专利技术开发新型微凝胶，用于将多肽A靶向大脑(专利编号62/362,105,2016)。 新颖性、重要性和影响力： 这个项目使用了从细胞模型到动物试验的各种技术来提高我们对氧化应激和细胞死亡的知识。这项研究是必要的，因为某些化合物和环境毒素导致细胞死亡的机制尚不清楚。此外，对神经保护的新见解将有助于推动更有效化合物的开发。该项目将开发的专利技术与用于增强定向脑传递的新型微凝胶的生产相结合，为加拿大提供了令人振奋的机会，进一步增加其对新的生物和工程知识和技术的贡献。

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.