Abstract：
This article describes a step-by-step process of lumbar intrathecal injection of Evans blue dye and AAV9-EGFP in adult (2-month-old) and neonatal (postnatal day 10) mice. Intrathecal injection is a clinically translatable technique that has already been extensively applied in humans. In mice, intrathecal injection is considered a challenging procedure that requires a trained and experienced researcher. For both adult and neonatal mice, lumbar intrathecal injection is directed into the L5-L6 intervertebral space. Intrathecally injected material enters the cerebrospinal fluid (CSF) within the intrathecal space from where it can directly access the central nervous system (CNS) parenchyma. Simultaneously, intrathecally injected material exits the CSF with pressure gradient and enters the endoneurial fluid and ultimately the peripheral nerves. While in the CSF, the injectable material also enters the bloodstream and systemic circulation through the arachnoid villi. A successful lumbar intrathecal injection results in adequate biodistribution of the injectable material in the CNS, PNS, and peripheral organs. When correctly applied, this technique is considered as minimally invasive and non-disruptive and can be used for the lumbar delivery of any solute. © 2024 Wiley Periodicals LLC. Basic Protocol 1: C57BL/6 adult and P10 mice lumbar intrathecal injection Basic Protocol 2: Tissue collection and preparation for evaluating Evans blue dye diffusion Basic Protocol 3: Tissue collection and preparation for immunohistochemistry staining Basic Protocol 4: Tissue collection and vector genome copy number analysis.
摘要：
本文介绍了成年（2月龄）和新生（出生后第10天）小鼠腰椎鞘内注射伊文思蓝染料和AAV9-EGFP的分步过程。鞘内注射是一种临床上可翻译的技术，已广泛应用于人类。在老鼠身上,鞘内注射被认为是一个具有挑战性的程序，需要训练有素和经验丰富的研究人员。对于成年和新生小鼠，腰椎鞘内注射直接进入L5-L6椎间隙。鞘内注射的材料进入鞘内空间内的脑脊液（CSF），从那里它可以直接进入中枢神经系统（CNS）实质。同时,鞘内注射的物质以压力梯度离开CSF，进入神经内膜液，最终进入周围神经。在脑脊液中，注射材料也通过蛛网膜绒毛进入血液和全身循环。成功的腰椎鞘内注射导致可注射材料在CNS中的充分生物分布。PNS,和外周器官。如果正确应用，该技术被认为是微创和非破坏性的，可用于任何溶质的腰椎输送。©2024Wiley期刊有限责任公司。基本方案1:C57BL/6成年小鼠和P10小鼠腰椎鞘内注射基本方案2:用于评估伊文思蓝染料扩散的组织收集和制备基本方案3:用于免疫组织化学染色的组织收集和制备基本方案4:组织收集和载体基因组拷贝数分析。
