Optical platform to image neuronal and vascular effects of cocaine in awake rodents

内侧前额叶皮层（mPFC）在可卡因成瘾中起着关键作用。的机制 可卡因滥用者的mPFC功能障碍尚未得到很好的理解，可能反映了脑血液的破坏 流动（可卡因的血管效应）和/或前额叶皮质多巴胺能调节的中断 （可卡因的神经效应）。然而，目前的神经成像工具不能容易地将血管与神经系统分离。 神经元效应也不能区分细胞特异性（例如，D1 r与D2 r神经元）对可卡因的反应。 这个应用程序，“光学平台，以图像神经元和血管的可卡因在清醒的啮齿动物的影响”， 建议开发一种创新的集成光学成像平台，以应对这些挑战 并证明其在成瘾研究中的价值。具体来说，我们建议开发一种独特的光学 该平台能够同时（1）对D1 r/D2 r特定神经元Ca 2+动力学进行荧光成像 （神经元活动的标志物）利用cre转基因小鼠和光学编码的 Ca 2+指示剂（GCaMP 6 f），以识别特定的神经元群体，和（2）三维超高分辨率光学 应用1.3 μ m的相干多普勒断层扫描技术（1.3 μ m ODT）对脑深部微血流（CBF）进行了研究。 mPFC（>1.4mm）。我们将证明这种综合方法评估血管和 急性和慢性可卡因清醒动物mPFC的神经元反应。 我们最近的光学工具进展及其用于研究可卡因在脑血管中的作用 D1 r和D2 r表达神经元的网络和神经元功能分别奠定了坚实的基础 为可卡因诱导mPFC功能障碍的拟议研究奠定基础。成功发展 提出的新的荧光-CONDODT平台不仅将提供新的见解可卡因诱导 mPFC功能障碍的潜在机制，但也将为研究神经血管相互作用的价值 以及它们在各种脑部疾病的动物模型中的破坏作用。

The medial prefrontal cortex (mPFC) plays a critical role in cocaine addiction. The mechanism underlying mPFC dysfunction in cocaine abusers are not well understood and could reflect disruption of cerebral blood flow (cocaine’s vascular effects) and/or disruption of dopaminergic modulation of the prefrontal cortex (cocaine’s neuronal effects). However, current neuroimaging tools cannot readily separate vascular from neuronal effects nor distinguish between cell specific (e.g., D1r vs D2r neuronal) responses to cocaine. This application, "Optical platform to image neuronal and vascular effects of cocaine in awake rodents", proposes to develop an innovative and integrated optical imaging platform to address these challenges and demonstrate its value in addiction research. Specifically, we propose to develop a unique optical platform that enables simultaneous (1) fluorescence imaging of D1r/D2r-specific neuronal Ca2+ dynamics (marker of neuronal activity) taking advantage of cre transgenic mice and viral delivery of optically encoded Ca2+ indicator (GCaMP6f) to identify specific neuronal populations, and (2) 3D ultrahigh-resolution optical coherence Doppler tomography (1.3m ODT) of micro cerebral blood flow (CBF) in deep layers of the mPFC (>1.4mm). We will demonstrate the value of this integrated approach to assess the vascular and neuronal responses in mPFC of awake animals to acute and chronic cocaine. Our recent optical tool advances and their use for studying the effects of cocaine in cerebrovascular networks and in neuronal function separately for D1r- and D2r- expressing neurons have laid a solid foundation for the proposed study of cocaine-induced mPFC dysfunction. Successful development of the proposed novel fluorescence-ODT platform will not only provide new insights into cocaine-induced mechanism underlying mPFC dysfunction but also will be of value for studying neurovascular interactions and their disruption in animal models of various brain disorders.