Cranial window implantation on mouse cortex to study microvascular change induced by cocaine
Cranial window implantation on mouse cortex to study microvascular change induced by cocaine
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DOI：
10.3978/j.issn.2223-4292.2014.11.31
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发表时间：
2015-02-01
影响因子：
2.8
通讯作者：
Pan, Yingtian
中科院分区：
文献类型：
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作者：
Park, Kicheon;You, Jiang;Pan, Yingtian
可卡因引起的中风是与可卡因滥用有关的最严重的医疗并发症之一。然而，在何种程度上，慢性可卡因可能会诱导沉默的微缺血诱发脑组织神经毒性尚未调查，部分原因是目前的神经成像工具的局限性，即缺乏高时空分辨率和灵敏度，同时测量脑血流量（CBF）在不同口径的血管定量和大视场（FOV）。光学相干断层扫描（OCT）技术使我们能够对脑血管网络（包括动脉、静脉和毛细血管）进行三维成像，并提供脑血管系统的高分辨率血管造影和网络中单个血管内的定量CBF速度（CBFv）。为了监测体内脑沿着慢性可卡因暴露的神经血管变化，我们开发了一种在小鼠脑上植入颅窗以实现长期皮质成像的方法。将颅窗植入两组动物的感觉运动皮质区，即，对照组[盐水处理，类似于0.1 cc/10 g/天，腹膜内注射（i. p.）]慢性可卡因组（可卡因治疗，30 mg/kg/d，i. p.）。植入后，通过OCT和立体镜定期对个体动物的皮质进行成像，以提供脑血管网的血管造影和定量CBFv，以及脑的表面成像。我们已经观察到血管血液动力学变化（即，CBFv变化），包括手术后不久脑的炎症反应（即，5天）在大脑中。重要的是，通过与对照动物进行比较，可以校准皮质中与药物相关的血管生理学变化，从而可以在可卡因动物中确定可卡因诱导的神经血管系统中的血液动力学变化。我们的研究结果表明，这种方法可以用来探讨神经血管功能的变化所引起的脑部疾病，如药物成瘾。
Cocaine-induced stroke is among the most serious medical complications associated with cocaine's abuse. However, the extent to which chronic cocaine may induce silent microischemia predisposing the cerebral tissue to neurotoxicity has not been investigated; in part, because of limitations of current neuroimaging tools, that is, lack of high spatiotemporal resolution and sensitivity to simultaneously measure cerebral blood flow (CBF) in vessels of different calibers quantitatively and over a large field of view (FOV). Optical coherence tomography (OCT) technique allows us to image three dimensional (3D) cerebrovascular network (including artery, vein, and capillary), and provides high resolution angiography of the cerebral vasculature and quantitative CBF velocity (CBFv) within the individual vessels in the network. In order to monitor the neurovascular changes from an in vivo brain along with the chronic cocaine exposure, we have developed an approach of implanting a cranial window on mouse brain to achieve long-term cortical imaging. The cranial window was implanted on sensorimotor cortex area in two animal groups, i.e., control group [saline treatment, similar to 0.1 cc/10 g/day, intraperitoneal injection (i.p.)] and chronic cocaine group (cocaine treatment, 30 mg/kg/day i.p.). After implantation, the cortex of individual animal was periodically imaged by OCT and stereoscope to provide angiography and quantitative CBFv of the cerebral vascular network, as well as the surface imaging of the brain. We have observed vascular hemodynamic changes (i.e., CBFv changes) induced by the cranial preparation in both animal groups, including the inflammatory response of brain shortly after the surgery (i.e., 5 days) in the brain. Importantly, by comparing with the control animals, the surgical-related vascular physiology changes in the cortex can be calibrated, so that the cocaine-induced hemodynamic changes in the neurovasculature can be determined in the cocaine animals. Our results demonstrate that this methodology can be used to explore the neurovascular functional changes induced by the brain diseases such as drug addiction.