鼠标显微注水技术进行有针对性的物质传递到中枢神经系统<i>通过</i>的颈内动脉
Summary
The present protocol describes a mouse microsurgery infusion technique, which effectively delivers substances directly into the brain via the internal carotid artery.
Abstract
Animal models of central nervous system (CNS) diseases and, consequently, blood-brain barrier disruption diseases, require the delivery of exogenous substances into the brain. These exogenous substances may induce injurious impact or constitute therapeutic strategy. The most common delivery methods of exogenous substances into the brain are based on systemic deliveries, such as subcutaneous or intravenous routes. Although commonly used, these approaches have several limitations, including low delivery efficacy into the brain. In contrast, surgical methods that locally deliver substances into the CNS are more specific and prevent the uptake of the exogenous substances by other organs. Several surgical methods for CNS delivery are available; however, they tend to be very traumatic. Here, we describe a mouse infusion microsurgery technique, which effectively delivers substances into the brain via the internal carotid artery, with minimal trauma and no interference with normal CNS functionality.
Introduction
在中枢神经系统中的体内模型（CNS）的疾病需要有效递送的外源物质，如药物，病原体，或外来体，进入大脑。因此，一个理想的递送方法应引起创伤小的动物，保护神经元网络的完整性，并在大脑1达到高的物质的浓度。
本地物质递送的若干外科手术方法已被描述，包括内护套，脑内，脑室内和注射或植入2，3，4，5。这些方法，但是，被认为是创伤至CNS，并且只允许感兴趣的物质的量低的施用。此外，已经提出，外源物质可以通过脑脊液6被快速移除</s向上>，和当采用上述技术的低渗透至大脑软组织已经观察到7。全身递送方法，如口服，肺部，皮下，和静脉内途径，在动物模型中比较常用的，尽管它们在物质递送至CNS表现出低效力，由于由其他器官8,9摄取。因此，分娩这些路线需要提升的剂量给药物质，增加副作用和毒性10，11的风险。
这里，我们描述了小鼠输注显微技术，其有效地提供了物质直接进入通过内部颈动脉的大脑。除了靶向递送至CNS，该技术不绕过正常的生理障碍，因此是biologica高度相关的涉及治疗剂或病原体进入大脑的通路升进程。
Protocol
Representative Results
Discussion
这里所描述的输液显微已被证明是在提供各种生物特征外源物质进入中枢神经系统，以防止整个身体1,12不需要的传播非常成功。血 – 脑屏障的破坏为几种中枢神经系统有关的疾病的病理学特征;因此评估外源物质与血脑屏障的关系是非常重要和兴趣。
提出这种手术模型导致有限创伤给动物，并且具有非常低的死亡率<sup…
Disclosures
The authors have nothing to disclose.
Acknowledgements
We would like to thank Dr. Lei Chen (Icahn School of Medicine at Mount Sinai, NY) who first established the use of this model in our laboratory, and to Dr. Gretchen Wolff (German Cancer Research Center, Heidelberg, Germany) for disseminating the technique in our laboratory. Supported in part by HL126559, DA039576, MH098891, MH63022, MH072567, DA027569, and NSC 2015/17/B/NZ7/02985.
Materials
| Anesthesia instrument | Vetequip | 901806 |
| Surgical scissors | Fine Science Tool | 14558-09 |
| Surgical forceps straight tip | Fine Science Tool | 00108-11 |
| Surgical forceps angled tip | Fine Science Tool | 00109-11 |
| Spring scissors | Fine Science Tool | 15000-08 |
| Nylon suture | Braintree Scientific | SUT-S 104 |
| Capillary tubing (Micro-Renathane 0.010” x 0.005” per ft.) | Braintree Scientific | MRE01050 |
| Closing suture | VWR | 95057-036 |
| Isoflurane | Piramal | |
| 2,3,5-Triphenyltetrazolium chloride | FisherScientific | 50-121-8005 |
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