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（D2 R）拮抗作用后行为超敏反应发展的分子机制，以及（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）使用活体大鼠脑成像研究氧化应激（OS）诱导的多巴胺转运蛋白和受体的变化，以及（4）通过开发新型微凝胶将肽A靶向递送至大脑来增强神经保护。目标和方法 * 短期5年：1。为了研究新型肽A对氟哌啶醇诱导的OS的神经保护作用，将用氟哌啶醇、氟哌啶醇+肽A和无活性肽B处理各组大鼠28天。将通过记录刻板行为、空咀嚼运动（VCM）和自发活动来监测行为超敏反应。2.为了确定肽A是否可以防止纹状体扁桃体神经元、纹状体管膜神经元和星形胶质细胞中的AIF移位，将用氟哌啶醇、氟哌啶醇+肽A和溶剂溶液处理各组大鼠28天。在治疗结束时，将解剖大脑区域并用神经元和星形胶质细胞的特异性抗体标记。3.为了确定是否存在谷氨酸释放的增加和GABA的减少，将如上文目的2中所述对大鼠进行处理。将通过微透析在活体大鼠中测量谷氨酸盐和GABA。4.为了研究肽A在细胞模型中的分子机制，D2 R转染的人神经母细胞瘤SH SY 5 Y细胞将用于测量自由基形成、MEF 2和Nrf 2表达以及用环境毒素、百草枯和氟哌啶醇处理后的细胞活力。5年后的长期目标：1。使用脑成像确定肽A的跨膜递送是否防止氧化应激期间多巴胺转运蛋白和D2 R的下调。2.使用我们的专利技术（专利号62/362，105，2016）开发新型微凝胶，用于将肽A靶向递送至大脑。*新奇，意义和影响：* 该计划使用从细胞模型到动物试验的技术，以提高我们对氧化应激和细胞死亡的认识。这项研究是必不可少的，因为某些化合物和环境毒素引起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。结合用于增强靶向脑递送的新型微凝胶的生产，将在该项目中开发的可专利技术为加拿大提供了令人兴奋的机会，以进一步增加其对新型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.***